WO2016072939A1 - Formulations comprenant des agents antimicrobiens avec des parties hydrophobes et leurs utilisations - Google Patents

Formulations comprenant des agents antimicrobiens avec des parties hydrophobes et leurs utilisations Download PDF

Info

Publication number
WO2016072939A1
WO2016072939A1 PCT/SG2015/050435 SG2015050435W WO2016072939A1 WO 2016072939 A1 WO2016072939 A1 WO 2016072939A1 SG 2015050435 W SG2015050435 W SG 2015050435W WO 2016072939 A1 WO2016072939 A1 WO 2016072939A1
Authority
WO
WIPO (PCT)
Prior art keywords
composition
cyclodextrin
hppcd
hydroxypropyl
nacl
Prior art date
Application number
PCT/SG2015/050435
Other languages
English (en)
Inventor
Chi Lui Paul HO
Chun Hwee Desmond TEO
Woon Pei Jeanette TEO
Hai Ning PEE
Original Assignee
National University Of Singapore
National University Hospital(S) Pte Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by National University Of Singapore, National University Hospital(S) Pte Ltd filed Critical National University Of Singapore
Publication of WO2016072939A1 publication Critical patent/WO2016072939A1/fr

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
    • A61K31/351—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom not condensed with another ring
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/075—Ethers or acetals
    • A61K31/085—Ethers or acetals having an ether linkage to aromatic ring nuclear carbon
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/4427—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems
    • A61K31/444—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems containing a six-membered ring with nitrogen as a ring heteroatom, e.g. amrinone
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
    • A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
    • A61K47/38—Cellulose; Derivatives thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
    • A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
    • A61K47/40—Cyclodextrins; Derivatives thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0043—Nose
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P33/00—Antiparasitic agents
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Definitions

  • the present invention relates to the field of microbiology.
  • the present invention relates to the use of a composition comprising antimicrobial agents for the prevention or treatment of infections, or for decolonization.
  • HAIs hospital-acquired, infections
  • Surgical site infections are the second most prevalent hospital- acquired infections in the United States with approximately 300,000 occurrences yearly, preceded only by urinary tract infections. Surgical site infections complicate about 5% of surgeries, where the severity ranges from superficial skin infections to life-threatening conditions.
  • Staphylococcus aureus SA is commonly isolated in surgical site infections, where most strains are methicillin -resistant (MRS A). The high costs and morbidities associated with surgical site infections have drawn great concern.
  • Intranasal mupirocin and photodisinfection have shown to reduce surgical site infections risks by a significant degree. However, their effectiveness may be limited by the anatomical restrictions of the nasal cavity, as the current methods have a finite capacity to reach the tortuous areas of the nose, especially the sinuses. For the application of ointments, for example, the extent of application is dependent on the skill of the caregiver; while for photodisinfection, it varies based on the flexibility and length of the probe used.
  • the present invention refers to a composition comprising one or more antimicrobial agent with hydrophobic moieties and a cyclodextrin.
  • the present invention refers to a composition as defined herein for use in treating an infection.
  • the present invention refers to a method of treating or preventing an infection in a patient, or for decolonizing an orifice of a patient, wherein the method comprised administering to the patient a therapeutically effective amount of the compositions as defined herein
  • the present invention refers to use of the composition as defined herein in the manufacture of a medicament for preventing or treating an infection, or for decolonizing an orifice of a patient.
  • the present invention refers to a method of producing the compositions as described herein, the method comprising: (a) mixing a ground cyclodextrin, as defined herein, and one or more ground antimicrobial agent, as defined herein; and (b) constantly agitating and heating the mixture formed under (a) .
  • Fig. 1 shows a chromatogram of mupirocin obtained with the mupirocin HPLC assay parameters as described in the experimental section and the example section below.
  • FIG. 2 shows further chromatograms depicting mupirocin undergoing acid degradation at 40°C. Indicated in arrows are peaks from the degraded products of mupirocin.
  • FIG. 3 shows further chromatograms depicting mupirocin undergoing base degradation at room temperature. Indicated in arrows are peaks from the degraded products of mupirocin.
  • Fig. 4 shows further chromatograms showing mupirocin undergoing base degradation at 40°C. Indicated in arrows are peaks from the degraded products of mupirocin.
  • Fig. 5 shows a line chart depicting the stability of MPC-HPpCD-NaCl in aqueous solution, evaluated over 8 weeks stored at room temperature with light exposure. The composition is considered to be stable when exposed to this condition.
  • Fig. 6 shows a line chart depicting the stability of MPC-HPpCD-NaCl in aqueous solution, evaluated over 8 weeks stored at room temperature without light exposure. The formulation is considered to be stable when exposed to this condition.
  • Fig. 7 shows a scatterplot depicting the increase in concentration of 2-hyrdoxypropyl-P-cyclodextrin (HPpCD) increases the solubility of mupirocin in water when reconstituted.
  • HPpCD 2-hyrdoxypropyl-P-cyclodextrin
  • Fig. 8 shows a column chart showing the concentration of mupirocin at saturation in 2-hydroxypropyl-P-cyclodextrin (HPpCD) and varying concentrations of carboxymethyl cellulose (CMC) and sodium chloride (NaCl).
  • CMC carboxymethyl cellulose
  • NaCl sodium chloride
  • FIG. 9 shows a photograph of a bacterial plate with a result of the time-dependent bactericidal activity for a mupirocin composition.
  • MRSA 2 was added to mupirocin- 2-hydroxypropyl-P-cyclodextrin (MPC-HPpCD) composition and the bacteria was plated at 15 minutes. This plate was observed after 24 hours incubation. The bacteria had been plated at dilutions of undiluted (ND), lxlO 1 (1E1) dilution, lxlO 3 (1E3) dilution and lxlO 2 (1E2) dilution (clockwise from left top).
  • Fig. 10 shows a line chart depicting the time-kill curves of mupirocin against MRSA 2 (A) and MRSA 7 (B).
  • Bactericidal activity is defined as a "reduction of 99.9% (> 3 logio) of the total number of CFU/mL in the original inoculum".
  • the bactericidal level is indicated by the dashed line.
  • the MRSA 2 bacteria strain is MPC resistant.
  • Fig. 11 shows a chromatogram of octenidine obtained with the octenidine HPLC assay parameters as described below.
  • Fig. 12 shows the Ql MS spectrum of OCT-HPpCD-NaCl dry powder formulation prepared two months prior to the formulation. Shown here is the total ion count (TIC) comparable to the freshly prepared formulation in Fig. 13 indicating stability of the sample.
  • Fig. 14 shows a scatterplot of data points depicting octenidine saturation in a composition comprising 2-hyrdoxypropyl-P-cyclodextrin (HPpCD) or a-cyclodextrin (a-CD).
  • HPpCD 2-hyrdoxypropyl-P-cyclodextrin
  • a-CD a-cyclodextrin
  • Fig. 15 shows a column chart showing that the increase in concentration of 2-hyrdoxypropyl-P-cyclodextrin (HPpCD) and a-cyclodextrin (a-CD) increases the solubility of octenidine in water in the presence of 2.8% sodium chloride (NaCl). It is observed that the concentration of octenidine dissolved in water in the presence of 2.8% NaCl, without cyclodextrin, is only 0.11 mg/ml.
  • HPpCD 2-hyrdoxypropyl-P-cyclodextrin
  • a-CD a-cyclodextrin
  • Fig. 16 shows a diagram of a 96-well, round-bottom, microtiter plate layout for Minimum Inhibitory Concentration (MIC) performed using the Broth Microdilution Method with 7 different strains of Staphylococcus aureus. The minimum inhibitory concentration was recorded as the lowest antimicrobial concentration where no bacteria growth is observed after overnight incubation.
  • MIC Minimum Inhibitory Concentration
  • Fig. 17 shows a photograph of a bacterial plate with a result of the time-dependent bactericidal activity for an octenidine composition.
  • MRSA 2 was added to OCT-HPpCD formulation and the bacteria were plated at 30 minutes. This plate was observed after 24 hours incubation. The bacteria were plated at dilutions of undiluted (ND), lxlO 1 (1E1) dilution, 1x10 3 (1E3) dilution and 1x102 (1E2) dilution (clockwise from left top). It is observed that there were no bacteria colonies in non-diluted and lxlO 1 (1E1) dilution quadrants due to the residual drug inhibiting the growth of surviving bacteria.
  • Fig. 18 shows a line chart depicting the time-kill curves of Octenidine against MRSA 7.
  • Bactericidal activity is defined as a "reduction of 99.9% (> 3 logio) of the total number of CFU/mL in the original inoculum". The bactericidal level is indicated by the dashed line.
  • Fig. 19 shows a chromatogram of triclosan obtained with the triclosan HPLC assay parameters.
  • Fig. 20 shows a scatterplot of data points depicting the solubility of triclosan in compositions produced using the non-aqueous (dry heat mixing) method as described herein.
  • the non-aqueous dry physical mixing method produces a dry powder complex for reconstitution.
  • HPpCD 2-hyrdoxypropyl-P-cyclodextrin
  • CMC carboxymethyl cellulose
  • TCS aqueous solubility of triclosan
  • Fig. 21 shows a scatterplot of data points depicting the solubility of triclosan in lyophilised compositions produced as described herein. Similar observations as shown in Fig. 18 above were seen for the aqueous method of slurry complexation. The slurry obtained from kneading is placed under a freeze-drying process to obtain a dry powder. The optimal composition is also at 40mM HPpCD and 1% w/v CMC, for a concentration of 0.3% w/v TCS.
  • Fig. 22 shows a line chart depicting the viscosities of MilliQ water (negative control/baseline) and the formulations containing concentrations of carboxymethyl cellulose (CMC) between 0.25% to 5% w/v CMC were measured. Concentrations up to 1% w/v CMC had comparable viscosities to MilliQ water, allowing for use in a nebuliser suitably.
  • CMC carboxymethyl cellulose
  • Fig. 23 shows the Ql MS Spectrum of TCS-HPpCD-NaCl dry powder formulation prepared two months prior to the formulation shown in Fig. 24. Shown here is the total ion count (TIC) comparable to the freshly prepared formulation in Fig. 24 indicating stability of the sample.
  • TIC total ion count
  • Fig. 25 shows a line plot depicting data showing the stability of aqueous TCS- HPpCD-NaCl formulation evaluated over 72 hours stored at room temperature with light exposure. The formulation is considered to be stable when exposed to this condition.
  • Fig. 26 shows a line plot depicting data showing the stability of aqueous TCS- HPpCD-NaCl formulation evaluated over 72 hours stored at room temperature without light exposure. The formulation is considered to be stable when exposed to this condition.
  • Fig. 27 is a photograph showing the full experimental set-up of the nebulising procedure for swab samples obtained from twenty healthy volunteers.
  • Fig. 28 is a photograph showing examples of experimental culture tubes before 72 hour incubation at 37°C. Negative control ("Control"), positive control (“CI 9”) and test (“T19”) are shown here.
  • Fig. 29 is a photograph showing examples of experimental culture tubes after 72 hour incubation at 37°C. Negative control ("Control"), positive control ("CI 3") and test (“T13") are shown here. Turbidity can be observed for the tube containing the positive control.
  • Fig. 30 is a photograph showing the side -by-side comparison of a positive control ("C13") and test ("T13") after 72 hour incubation at 37°C. Turbidity can be observed for the positive control, while a clear solution is seen for the test.
  • Fig. 31 shows a line chart depicting the time-kill curves of triclosan against MRS A 2 (A) and MRSA 7 (B).
  • Bactericidal activity is defined as a "reduction of 99.9% (> 3 logio) of the total number of CFU/mL in the original inoculum". The bactericidal level is indicated by the dashed line.
  • HAIs hospital-acquired infections
  • a greater percentage of people who are hospitalized today are likely to be seriously ill with more weakened immune systems than in the past.
  • some medical procedures bypass the body's natural protective barriers. Since medical staff routinely moves from patient to patient, the staff themselves serve as a means for spreading pathogens, thus becoming possible vectors for the transmission of such infections.
  • Microbes found in these environments can more often than not survive for a long time on surfaces in the hospital and enter the body through, for example, wounds, catheters, and ventilators. Orifices in the body of a patient can thus become susceptible to and can even facilitate various infections in a hospital setting, especially where the immune system is weak or when the patient becomes exposed to resistant microbes.
  • compositions used for such purposes and which are known in the art are inferior solubility of active substances, such as antimicrobial agents with hydrophobic moieties or hydrophobic antimicrobial agents, resulting in formulations which are, for example, not easily applicable.
  • compositions comprising an antimicrobial agent and a cyclodextrin.
  • composition and “formulation” are used herein are interchangeable.
  • the composition comprises an antimicrobial agent with hydrophobic moieties and a cyclodextrin.
  • the antimicrobial agent is hydrophobic.
  • hydrophobic refers to a physical property of any molecule to seemingly repelled from a mass of water, that is to say, there is a lack of attractive forces between the molecule and the solvent, in this case water.
  • hydrophobic moiety refers to the physical property of a portion of a molecule to lack an affinity to water.
  • hydrophobic moieties may also contain hydrophilic moieties and may also display the same characteristics as a hydrophobic molecule.
  • a hydrophobic molecule is understood to be hydrophobic in all moieties. This hydrophobicity of the antimicrobial agent also plays a role in its ability to be dissolved in water. It is understood in the art that hydrophobic compounds are or compounds with hydrophobic moieties may be difficult to dissolve, that is the solubility of hydrophobic compounds is low, whereas hydrophilic compounds are easily dissolved in water.
  • an antimicrobial agent refers to a hydrophobic agent or agents that are able to complex with cyclodextrins or their derivatives used to kill or inhibit the growth of microbes and microorganisms.
  • This agent may be chemical or biological in nature, or, synthetic or natural in origin.
  • An antimicrobial agent may include, but is not limited to, antibacterial agents, antiseptic agents, antibiotics, fungicides, bacteriostats, sanitisers, disinfectants and sterilisers.
  • an antimicrobial agent may be, but are not limited to ethyl alcohol, isopropyl alcohol, benzalkonium chloride, cetrimide, methylbenzethonium chloride, benzethonium chloride, cetalkonium chloride, cetylpyridinium chloride, dofanium chloride, domiphen bromide, chlorhexidine gluconate, chlorhexidine acetate, proflavine hemisulphate, triphenylmethane, brilliant green, crystal violet, gentian violet, hydrogen peroxide solution, potassium permanganate solution, benzoyl peroxide, chlorocresol, chloroxylenol, chlorophene, hexachlorophane/hexachlorphene, mupirocin calcium, octenidine dihydrochloride, triclosan, polyhexanide, chlorhexidine triclocarban ,hydroxyquinoline sulphate, potassium hydroxyquinoline sulphate, chlorquinaldol
  • the antimicrobial agent may be triclosan, octenidine dihydrochloride, mupirocin calcium or derivatives thereof.
  • the antimicrobial agent is mupirocin (also mupirocin calcium: MPC).
  • the antimicrobial agent is octenidine (also octenidine dihydrochloride; OCT).
  • the antimicrobial agent is triclosan (TCS).
  • Combinations of various antimicrobial agents may also be used. These combinations may comprise at least one or more antimicrobial agents, for example two, three or more antimicrobial agents.
  • triclosan may alternatively be used as a preservative.
  • the composition may be formulated with, for example, mupirocin, to overcome resistance to some mupirocin-resistant MRSA. Therefore, in one example, the composition may comprise mupirocin and triclosan. In another example, the composition may comprise mupirocin and octenidine. In yet another example, the composition may comprise octenidine and triclosan.
  • the bispyridinamine octenidine dihydrochloride is a cationic compound with two positively-charged active centers together with long hydrocarbon chains. It is a broad spectrum antimicrobial agent, effective against both Gram-negative and Gram-positive bacteria including MRSA, as well as certain fungi. This is due to its ability to adhere and interact with the cell wall and cell wall components. Aside from its wide spectrum of activity, octenidine dihydrochloride has generally superior antimicrobial efficacy in vitro and bactericidal activity as compared to chlorhexidine and alexidine. It is also proven to be more effective than chlorhexidine and alexidine in the inhibition of plaque-forming enzymes of Streptococcus mutans leading to better oral health.
  • octenidine dihydrochloride can also be formulated as a mouthwash.
  • Octenidine is not absorbed during oral or topical administration. Its ability to bind readily to negatively-charged surfaces allows it to exert a long residual effect. Furthermore, bacteria are unlikely to develop resistance to octenidine dihydrochloride due to its mechanism of action. There is also no difference in its effectiveness against MRSA and MSSA bacteria strains.
  • the antimicrobial agent is octenidine or a derivative thereof. In another example, the antimicrobial agent is octenidine dihydrochloride.
  • Formulations of octenidine dihydrochloride with and without hypertonic saline can have applications in nasal decolonisation, reducing the risk of surgery-related infections.
  • octenidine dihydrochloride compositions with hypertonic saline can be used as an adjunctive treatment for many sino-nasal conditions such as sinusitis, rhinitis and respiratory diseases such as bronchiolitis.
  • a saline concentration of more than 3.0% could cause irritation to the mucous membrane, a concentration of 2.8% sodium chloride was adopted during formulation development.
  • the nasal spray, irrigation or nebulised solution of octenidine dihydrochloride formulation with or without sodium chloride may also be administered before an operation for nasal decolonization.
  • a formulation of the compositions as described herein comprising an antimicrobial agent is considered for nasal decolonisation.
  • Mupirocin is an antibiotic produced by the bacterium Pseudomonas fluorescens which acts on isoleucyl tRNA synthetase and thereby inhibiting protein synthesis, eventually leading to the bacteria's death. It has activity against most gram-positive and gram-negative bacteria.
  • mupirocin is used as an ointment at 2% concentration.
  • the standard of case is application of mupirocin 2% ointment two to three times daily, for up to ten days.
  • the nasal carriage is generally decolonized after five to seven days of treatment.
  • the present disclosure refers to a composition comprising an antimicrobial agent, wherein the antimicrobial agent is mupirocin or a derivative thereof.
  • the antimicrobial agent is mupirocin calcium.
  • Triclosan is an antibacterial and antifungal agent which is shown to have broad spectrum activity, and is particularly effective against Methicillin-resistant Staphylococcus aureus (MRSA).
  • MRSA Methicillin-resistant Staphylococcus aureus
  • triclosan acts as a biocide with multiple cytoplasmic and membrane targets.
  • triclosan appears bacteriostatic, and it targets bacteria primarily by inhibiting fatty acid synthesis. Use of triclosan in consumer products is prevalent.
  • compositions as described herein comprise the antimicrobial agent triclosan or a derivative thereof.
  • Antimicrobial agents may be effective at different concentrations, depending on the microorganism in question and the desired effect, that is, if the intention is to eradicate or to impede growth and/or proliferation of the microorganism.
  • the concentration of the antimicrobial agent in the composition may be between about 0.1% w/v to about 10% w/v, between about 0.05% w/v to about 5% w/v, between about 0.1% w/v to about 4% w/v, between about 0.3% w/v to about 2% w/v, between about 0.1% w/v to about 1.5% w/v, between about 0.15% w/v to about 2% w/v, between about 2% w/v to about 3% w/v, between about 0.1% w/v to about 2% w/v, about 0.05% w/v, about 0.1% w/v, about 0.15% w/v, about 0.2% w/v, about 0.25% w/v,
  • the concentration of the antimicrobial agent in the composition is between about 0.1% w/v and about 2% w/v. In one example, the concentration of the antimicrobial agent in the composition is 0.1% w/v. In another example, the concentration of the antimicrobial agent in the composition is 0.3% w/v. In another example, the concentration of the antimicrobial agent in the composition is 2% w/v. As used herein, the term "w/v" refers to the ratio of weight per volume.
  • cyclodextrin (CD) or “cyclodextrins” refers to cyclic oligosaccharides that contain 6 or more D-(+) glucopyranose units that are attached by ⁇ , ⁇ , or ⁇ -(1 ,4) glucosidic bonds. It has been shown that cyclodextrins are able to form complexes with a variety of hydrophobic molecules due to their unique structure. Cyclodextrin derivatives are extensively used in research labs, for example to remove cholesterol from cultured cells and they are well known in the pharmaceutical industry for their ability to solubilise drugs.
  • Cyclodextrins are able to form inclusion complexes with, for example, many drugs by taking up the whole drug, or, more commonly, the lipophilic moiety of the drug molecule, into a cyclodextrin cavity.
  • cyclodextrins can be, but are not limited to, a-cyclodextrin (a-CD), ⁇ -cyclodextrin ( ⁇ -CD) and ⁇ -cyclodextrin ( ⁇ -CD), each containing six, seven, and eight glucopyranose units, respectively.
  • ⁇ -cyclodextrin is the most useful pharmaceutical complexing agent due to its cavity size, availability, low cost and other properties.
  • cyclodextrin is used in the compositions to improve the aqueous solubility of the antimicrobial agent in aqueous solution.
  • cyclodextrin is used to improve the aqueous solubility of triclosan (TCS), which is known as a poorly water-soluble phenolic antiseptic.
  • TCS triclosan
  • the composition therefore comprises triclosan and cyclodextrin.
  • cyclodextrin is used to improve the aqueous solubility of mupirocin, the composition thus comprising mupirocin and cyclodextrin.
  • cyclodextrin is used to improve the aqueous solubility of octenidine dihydrochloride, the composition thus comprising octenidine dihydrochloride and cyclodextrin.
  • a cyclodextrin or derivatives thereof may be, but are not limited to, hydroxypropyl- ⁇ - cyclodextrins and methylated cyclodextrins.
  • the cyclodextrin is ⁇ - cyclodextrin.
  • 2-hydroxypropyl ⁇ -cyclodextrin ( ⁇ ) is known to have a low toxicity as compared to other cyclodextrins. Therefore, in another example, the cyclodextrin is 2-hydroxypropyl ⁇ -cyclodextrin ( ⁇ ).
  • octenidine dihydrochloride has comparatively good solubility in water, it is unable to co-dissolve with 2.8% w/v sodium chloride (NaCl) and will precipitate in saline solution.
  • the formulation of octenidine dihydrochloride together with 2-hydroxypropyl ⁇ -cyclodextrin ( ⁇ ) greatly improved the aqueous solubility of octenidine dihydrochlroide in water in the presence of sodium chloride (NaCl).
  • This formulation when prepared with the addition of 2.8% w/v sodium chloride, improves the antiseptic and irrigative properties of the formulation.
  • the composition of the formulation comprises 0.1% w/v octenidine dihydrochloride (OCT) and 50 mM 2-hydroxypropyl ⁇ -cyclodextrin ( ⁇ ). 2.8% w/v sodium chloride can be added when required.
  • OCT-HPpCD and OCT- HPpCD-NaCl The pH of the octenidine dihydrochloride compositions (OCT-HPpCD and OCT- HPpCD-NaCl) is between 6 and 7, making it compatible with the nasal cavity which has an average pH of approximately 6.3.
  • the concentration of cyclodextrin required to improve the aqueous solubility of a antimicrobial agent may vary depending on the physic-chemical characteristics of the antimicrobial agent, such as chemical polarity, hydrophobicity, solubility, temperature, pressure, the solvent and the presence of other solutes in the solvent.
  • the cyclodextrin may be present in a range between about 20 mM to about 200 mM, between about 20 mM to 100 mM, between about 20 mM to about 30 mM, between about 25 mM to about 45 mM, between about 35 mM to about 50 mM, between about 55 mM to about 70 mM, between about 65 mM to about 85 mM, between about 80 mM to about 90 mM, between about 90 mM to about 100 mM, between about 85 mM to about 95 mM, or may be present in amount of about 20 mM, about 30 mM, about 35 mM, about 40 mM, about 45 mM, about 49 mM, about 50 mM, about 55 mM, about 60 mM, about 70 mM, about 80 mM, about 90 mM, about 95 mM, about 100 mM, about 110 mM, or about 200mM.
  • cyclodextrin is present in a concentration of about 40 mM to about 100 mM. In another example, cyclodextrin is present in a concentration of about 40 mM. In yet another example, cyclodextrin is present in a concentration of about 50 mM. In a further example, cyclodextrin is present in a concentration of about 100 mM.
  • polymers may be used to further enhance the aqueous solubility, at the same time allowing for lesser amounts of cyclodextrin required.
  • the polymers may also enhance the viscosity of the formulation to confer better adherence to the applied surface.
  • the composition may further comprise a polymer.
  • the polymer is absent.
  • the term "polymer" refers to a large molecule, or macromolecule, composed of many repeated subunits.
  • the polymer can be pharmaceutically or medically acceptable, especially for compositions intended for external, as well as internal use on a patient. Therefore, the polymer should not be toxic and be well soluble in aqueous solution, and should not compromise the solubility of the compound if it is used solely as a viscosity enhancer.
  • the concentration of the polymer present in the composition also influences the viscosity of the composition.
  • increasing the viscosity of a composition may increase the local bioavailability of the active compounds, for example in topical applications.
  • the polymer may be present in the composition in an amount between about 0.1% w/v to about 2% w/v, about 0.1% w/v to about 0.5% w/v, about 0.4% w/v to about 0.8% w/v, about 0.5% w/v to about 1% w/v, about 0.2% w/v to about 0.3% w/v, about 0.25% w/v to about 0.3% w/v or in an amount of about 0.1% w/v, about 0.18% w/v, about 0.20% w/v, about 0.23% w/v, about 0.24% w/v, about 0.25% w/v, about 0.3% w/v, about 0.8% w/v, about 0.9% w/v or about 0.12% w/v.
  • the polymer is present in an amount of between about 0.1% w/v to 2% w/v. In another example, the polymer is present in an amount of about 0.25% w/v. In yet another example, the polymer is present in an amount of about 1% w/v.
  • Polymers used for this purpose can include, but are not limited to cellulose derived polymers, such as salt derivatives of carboxymethyl cellulose (CMC), microcrystalline cellulose and derivatives thereof; soluble cellulose derivatives, such as hydroxypropyl cellulose, hydroxypropyl methylcellulose, hydroxypropyl cellulose (HPC), methyl cellulose, and derivatives thereof, xanthan gum; insoluble cellulose derivatives, such as ethylcellulose, microcrystalline cellulose (MCC), and derivatives thereof; polyacrylates, such as carbomers, polycarbophil, and derivatives thereof, polyvinyl alcohol; starch, chitosan and derivatives thereof, alginates, acacia, and combinations thereof.
  • CMC carboxymethyl cellulose
  • HPC hydroxypropyl cellulose
  • MMC microcrystalline cellulose
  • polyacrylates such as carbomers, polycarbophil, and derivatives thereof, polyvinyl alcohol
  • starch, chitosan and derivatives thereof alginates, aca
  • carboxymethyl cellulose sodium also increases the viscosity of the formulation, allowing for better adhesion, for example to the mucosal surfaces of the nasal passage, thus facilitating drug delivery. Therefore, in another example, the composition may further comprise a cellulose derivative, such as carboxymethyl cellulose or carboxymethyl cellulose sodium.
  • compositions as described herein may also further comprise a salt.
  • a salt may be used to improve tonicity of the resulting composition and therefore enable an improved pharmaceutical, medical or non-medical application.
  • tonicity refers to is a measure of the effective osmotic pressure gradient (as defined by the water potential of two solutions) of two solutions separated by a semipermeable membrane.
  • tonicity is the relative concentration of solutions that determine the direction and extent of diffusion. It is commonly used when describing the response of cells immersed in an external solution.
  • tonicity is influenced only by solutes that cannot cross the membrane, as only these exert an effective osmotic pressure. Solutes able to freely cross the membrane do not affect tonicity because they will always be in equal concentrations on both sides of the membrane.
  • the salt may be provided in the form of a crystalline compound, a partially crystalline compound or as a solution.
  • a salt such as sodium chloride (NaCl)
  • NaCl sodium chloride
  • saline solution refers to a sterile or non-sterile solution of sodium chloride (NaCl) in water.
  • Sodium chloride is generally used, particularly if a buffer containing sodium ions is used in the composition, and is typically present in an amount that is physiologically equivalent to the tonicity of the nasal membranes.
  • the saline solution may be a hypertonic, an isotonic or a hypotonic solution depending on the intended application.
  • the term “saline” can be used interchangeably with the term “salt water solution”.
  • Clinical trials indicate that use of a hypertonic saline can enhance the ciliary beating frequency (CBF), which provides a further benefit to the patients.
  • CBF ciliary beating frequency
  • Randomised controlled trials found that for patients with sinusitis, daily nasal irrigation with hypertonic saline was able to improve sinus-related quality of life and reduced the severity of symptoms.
  • Another randomised controlled trial conducted in patients with symptomatic allergic rhinitis similarly reported improved nasal clearance time and reduced nasal symptoms, which was defined as nasal obstruction, nasal itching, nasal discharge and sneezing.
  • Treatments using hypertonic saline are concluded to be more efficacious than normal saline (0.9% w/v sodium chloride) treatments.
  • Hypertonic saline may be formulated with octenidine dihydrochloride-2-hydroxypropyl-P-cyclodextrin (OCT- HPPCD), mupirocin calcium-2-hydroxypropyl-P-cyclodextrin (MPC-HPpCD) or triclosan-2- hydroxypropyl-P-cyclodextrin-carboxymethyl cellulose (TCS-HPpCD-CMC), respectively in aqueous solution.
  • Hypertonic saline nasal irrigation is widely regarded to be beneficial in supporting the mechanical clearance of mucus.
  • hypertonic saline can facilitate reduction of mucosal edema, enhance ciliary beat activity, decrease inflammation and removal of antigen, leading to a protective effect on sino-nasal mucosa.
  • Hypertonic saline is indicated as adjunctive treatment in many sino-nasal conditions. These include acute sinusitis, chronic sinusitis, allergic rhinitis, non-allergic rhinitis, atrophic rhinitis, sinonasal sarcoid, post-operative care and other scab-forming conditions.
  • the concentration of the salt present in the compositions described herein may vary based on the required application. Therefore, the salt present in the composition may be in an amount between about 2% w/v to about 4% w/v, between about 2% w/v to about 3% w/v, between about 2.5% w/v to about 3.5% w/v, between about 3.5% w/v to about 4% w/v, between about 2.75% w/v to about 3.15% w/v or in an amount of about 1.8% w/v, about 2% w/v, about 2.2% w/v, about 2.4% w/v, about 2.6 % w/v, about 2.8% w/v, about 3.0% w/v or about 3.2% w/v.
  • the salt is present in the composition at an amount of about 2.8% w/v. In another example, the salt is present in the composition at an amount of about 3% w/v. In another example, the salt may be absent. In one example, the salt is sodium chloride.
  • both 2-hydroxypropyl-P-cyclodextrin (HPpCD) and carboxymethyl cellulose (CMC) serve to improve the aqueous solubility of the formulation, at the same time allowing for lesser amounts of HPpCD required, and modulate the release of triclosan in the nasal cavity when nebulised.
  • CMC when present, also increases the viscosity of the formulation, allowing for better adhesion to the mucosal surfaces of the nasal passage, facilitating drug delivery.
  • Saline when added, is added for its bacteriostatic and irrigative properties, and also enhances the aqueous solubility of the formulation.
  • the present disclosure refers to a composition comprising triclosan (TCS), carboxymethyl cellulose (CMC) and 2-hydroxypropyl-P-cyclodextrin (HPpCD).
  • the composition may further comprise a salt, whereby the composition then comprises triclosan (TCS), carboxymethyl cellulose (CMC), 2-hydroxypropyl-P-cyclodextrin (HPPCD) and sodium chloride (NaCl).
  • the present disclosure refers to a composition
  • a composition comprising about 0.2% w/v to about 0.5% w/v triclosan (TCS), about 0.5% w/v to about 1.5% w/v carboxymethyl cellulose (CMC) and about 30 mM to about 50 mM 2 hydroxypropyl-P-cyclodextrin (HPpCD).
  • TCS w/v to about 0.5% w/v triclosan
  • CMC carboxymethyl cellulose
  • HPpCD hydroxypropyl-P-cyclodextrin
  • the composition comprises about 0.2% w/v to about 0.5% w/v triclosan (TCS), about 0.5% w/v to about 1.5% w/v carboxymethyl cellulose (CMC), about 30 niM to about 50 niM 2-hydroxypropyl-P- cyclodextrin (HPpCD) and about 0.5% w/v to about 3% w/v sodium chloride (NaCl). More precisely, in one example, a composition comprises 0.3% w/v triclosan (TCS), 1% w/v carboxymethyl cellulose (CMC) and 40 mM 2-hydroxypropyl-P-cyclodextrin (HPpCD).
  • TCS 0.3% w/v triclosan
  • CMC carboxymethyl cellulose
  • HPpCD 2-hydroxypropyl-P-cyclodextrin
  • the present disclosure refers to a composition
  • a composition comprising 0.3% w/v triclosan (TCS), 1% w/v carboxymethyl cellulose (CMC), 40 mM 2-hydroxypropyl-P-cyclodextrin (HPPCD) and 2.8% w/v sodium chloride.
  • HPpCD is used to improve the aqueous solubility of TCS, a poorly water-soluble phenolic antiseptic.
  • the composition of the triclosan-2-hydroxypropyl-P-cyclodextrin- carboxymethyl cellulose complex is 0.3% w/v triclosan, 40mM 2-hydroxypropyl-P-cyclodextrin (HPpCD), 1% w/v carboxymethyl cellulose (CMC).
  • Concentrations above 1% carboxymethyl cellulose improved the aqueous solubility of the formulation further but also increased the viscosity of the solution formed, making it less suitable for nebulisation.
  • 1% w/v saline can also be added to the formulation to improve the antiseptic and irrigative properties of the formulation.
  • the formulation can be combined with a hypertonic saline solution for the nebulisation process, which may be used as an adjunctive treatment for many sino-nasal conditions such as sinusitis, rhinitis.
  • This composition is for nasal decolonisation, that is, to reduce the risks of surgical site infections in patients undergoing surgery.
  • the formulation may also have secondary uses such as for nasal irrigation, nasal cleaning, or as an antiseptic. Formulation of the antiseptic solution is also considered.
  • the pH of the triclosan-based compositions ranges between about 6.43 to about 6.70.
  • the pH of the triclosan -based compositions ranges between about 6.72 to about 6.96.
  • the osmolality of an antimicrobial solution as described herein containing triclosan, without saline is between about 309 to about 320 mmol/kg and between about 387 to about 392 mmol/kg for the compositions with saline added.
  • the present disclosure refers to a composition comprising octenidine dihydrochloride (OCT) and 2-hydroxypropyl-P-cyclodextrin (HPpCD).
  • OCT octenidine dihydrochloride
  • HPpCD 2-hydroxypropyl-P-cyclodextrin
  • the composition comprises about 0.05% w/v to about 0.15% w/v octenidine dihydrochloride (OCT) and about 40 mM to about 60 mM 2-hydroxypropyl- ⁇ -cyclodextrin (HPpCD). More precisely, in one example, a composition comprises 0.1% w/v octenidine dihydrochloride (OCT) and 50 mM 2-hydroxypropyl-P-cyclodextrin (FfPpCD).
  • compositions described herein may further comprise a salt. Therefore, in a further example, a composition comprises octenidine dihydrochloride (OCT), 2-hydroxypropyl-P- cyclodextrin (HPpCD) and sodium chloride (NaCl). In yet another example, the composition comprises about 0.05% w/v to about 0.15% w/v octenidine dihydrochloride (OCT), about 40 mM to about 60 mM 2-hydroxypropyl-P-cyclodextrin (HPpCD) and about 0.5% w/v to 3% w/v sodium chloride (NaCl).
  • OCT octenidine dihydrochloride
  • HPpCD 2-hydroxypropyl-P-cyclodextrin
  • NaCl sodium chloride
  • the present disclosure refers to a composition comprising 0.1% w/v octenidine dihydrochloride (OCT), 50 mM 2-hydroxypropyl-P-cyclodextrin (HPpCD) and 2.8% w/v sodium chloride (NaCl).
  • OCT octenidine dihydrochloride
  • HPpCD 2-hydroxypropyl-P-cyclodextrin
  • NaCl sodium chloride
  • mupirocin MPC
  • HPpCD 2-hydroxypropyl- ⁇ -cyclodextrin
  • This formulation can also be prepared with the addition of 2.8% w/v sodium chloride (NaCl) to improve the antiseptic and irrigative properties of the formulation.
  • a composition comprising mupirocin comprises of 2% w/v mupirocin (MPC) and 100 mM 2-hydroxypropyl-P-cyclodextrin (HPpCD). 2.8% w/v sodium chloride (NaCl) may be added when required.
  • the pH of the MPC compositions (MPC-HPpCD and MPC-HPpCD-NaCl) is between 6 and 7, making it compatible with the nasal cavity which has an average pH of approximately 6.3.
  • the present disclosure refers to a composition comprising mupirocin calcium (MPC) and 2-hydroxypropyl-P-cyclodextrin (HPpCD).
  • the composition comprises about 1% w/v to 3% w/v mupirocin calcium (MPC) and about 50 mM to about 150 mM 2-hydroxypropyl-P-cyclodextrin (HPpCD). More precisely, in another example, the composition comprises 2% w/v mupirocin calcium (MPC) and 100 mM 2-hydroxypropyl-P-cyclodextrin (HPpCD).
  • the composition comprises mupirocin calcium (MPC), 2-hydroxypropyl-P-cyclodextrin (HPpCD) and sodium chloride (NaCl).
  • a composition comprises about 1% w/v to 3% w/v mupirocin calcium (MPC), about 50 mM to about 150 mM 2-hydroxypropyl-P-cyclodextrin (HPPCD) and about 0.5% w/v to 3% w/v sodium chloride (NaCl).
  • the composition comprises 2% w/v mupirocin calcium (MPC), 100 mM 2- hydroxypropyl-P-cyclodextrin (HPpCD) and 2.8% w/v sodium chloride (NaCl).
  • the compositions disclosed herein comprise carboxymethyl cellulose (CMC).
  • described herein is a composition which further comprises about 0.1% w/v to about 0.3% w/v carboxymethyl cellulose (CMC).
  • the present composition may also contain various pharmaceutically acceptable additives such as tolerance enhancers (sometimes more specifically referred to as humectants), absorption enhancers (sometimes also referred to as surfactants), preservatives, viscosity modifying agents (e.g., thickening agents), osmolality adjusters, complexing agents, stabilizers, solubilizers, or any combination thereof.
  • tolerance enhancers sometimes more specifically referred to as humectants
  • absorption enhancers sometimes also referred to as surfactants
  • preservatives e.g., viscosity modifying agents (e.g., thickening agents), osmolality adjusters, complexing agents, stabilizers, solubilizers, or any combination thereof.
  • a tolerance enhancer may be used in order to inhibit drying of the nasal membrane or mucosa.
  • a tolerance enhancer may also serve the purpose of inhibiting or relieving irritation of the nasal membranes.
  • suitable tolerance enhancers include, for example, humectants such as sorbitol, propylene glycol, glycerol, glycerin, hyaluronan, aloe, mineral oil, vegetable oil, soothing agents, membrane conditioners, sweeteners, and mixtures thereof.
  • the selection and concentration of a tolerance enhancer may depend on a number of factors, including, for example, the type and concentration of antimicrobial agent being used in the composition. When used, the concentration of the tolerance enhancer in the composition will typically be in amounts from about 0.01% w/w to about 20% w/w.
  • a preservative may also be employed to increase the shelf -life of the composition.
  • a number of well-known and pharmaceutically acceptable preservatives may be used in the present composition, including, for example, parabens, thimerosal, phenylcarbinol, chlorobutanol, benzalkonium chloride, or benzyl alcohol and combinations thereof.
  • Other ingredients which extend shelf life can be added such as for example, antioxidants.
  • antioxidants include sodium metabisulfite, potassium metabisulfite, ascorbyl palmitate and other pharmaceutically acceptable antioxidants.
  • the antioxidant will be present in the composition in a concentration of from about 0.01% w/w to about 5% w/w.
  • a surfactant or absorption enhancer may also be used in the composition in order to enhance the absorption of the antimicrobial compound across the nasal membrane.
  • the active agent is intended for topical use or dermal application, no such surfactant is added, as the active ingredient in not intended for internalization in a patient, that is, the active ingredient is not intended to pass the nasal mucosal membrane.
  • suitable absorption enhancers include non-ionic, anionic and cationic surfactants. Any of a number of well-known surfactants may be used, including, for example, polyoxyethylene derivatives of fatty acids, partial esters of sorbitol anhydrides, sodium lauryl sulfate, sodium salicylate, oleic acid, lecithin, dehydrated alcohol, Tween (e.g., Tween 20, Tween 40, Tween 60, Tween 80 and the like), Span (e.g., Span 20, Span 40, Span 80 and the like), polyoxyl 40 stearate, polyoxy ethylene 50 stearate, edetate disodium, propylene glycol, glycerol monooleate, fusieates, bile salts, octoxynol and combinations thereof.
  • surfactants e.g., polyoxyethylene derivatives of fatty acids, partial esters of sorbitol anhydrides, sodium lauryl s
  • the concentration of the surfactant in the composition will typically be from about 0.1% w/w to about 50% w/w.
  • concentrations of sodium salicylate, sodium lauryl sulfate and edetate disodium may be from about 0.01% to about 5% w/w of the composition.
  • Concentrations of polyoxyl 40 stearate, lecithin, dehydrated alcohol can be from about 0.1% to about 10% w/w of the composition.
  • Concentrations of oleic acid can be from about 0.01% to about 5% w/w of the composition.
  • Concentrations of propylene glycol and Tween 20 can be from about 0.1% to about 25% w/w of the composition.
  • a sweetening agent may also be added to the composition as described herein.
  • any one of the components may result in a bitter or otherwise displeasing taste, it would be possible to ameliorate this bitter taste by including an acceptable sweetening agent to the formulation.
  • This may be of use in formulations intended for oral application, nasal irrigation or as nasal sprays.
  • sweetening agents can be, but are not limited to, sugars such as monosaccharides, disaccharides and polysaccharides.
  • suitable sugars include but are not limited to xylose, ribose, glucose, mannose, galactose, fructose, dextrose, sucrose, sucralose, maltose, partially hydrolyzed starch or corn syrup solids, stevia, and sugar alcohols such as sorbitol, xylitol, mannitol, glycerin and combination thereof. If the addition of such a sweetening agent is contemplated, the amount added would need to be sufficient in order to mask the bitter or otherwise displeasing taste of the composition, but without impeding the efficacy of the resulting composition in its intended purpose.
  • the sweetening agent may, for example, be added together with a preservative as defined herein.
  • the amount of the sweetening agent present in the formulation may be between about 1% to about 40%, between about 0.01% to about 0.1%, between about 0.05% to about 0.5%, between about 1% to about 5%, between about 4% to about 10%, between about 8% to about 15%, between about 12% to about 15%, between about 15% to about 20%, between about 20% to about 35%, between about 25% to about 40%, about 8%, about 10%, about 15%, about 20%, about 30% or about 40%.
  • a sweetening agent in a formulation may, for example, result in the composition comprising 100 parts of water, 10 parts of xylitol/xylose, 0.65 parts of sodium chloride and effective amounts of benzalkonium chloride and phenylcarbinol as preservatives.
  • compositions for treating or preventing infections whereby the infection may be, but is not limited to, an ear infection, a nose infection, a throat infection, a sino-nasal infection, a sinus infection, a respiratory infection, rhinitis and bronchiolitis.
  • the composition of the invention may also be used as an adjunctive treatment for many sino-nasal conditions such as sinusitis, rhinitis.
  • the composition described herein is administered to a patient before or after surgery. Administration may also take place both before and after surgery.
  • compositions for treating or preventing infection in a patient refers to the invasion of a living organism by disease-causing microorganisms.
  • microorganism may be used interchangeably with "microbe”.
  • the microorganisms referred to herein may be pathogenic and non-pathogenic, as well as prokaryotic and eukaryotic microorganisms, such as, but not limited to bacteria, fungi, mould, protozoan and yeast.
  • the bacteria may be, but is not limited to, Salmonella spp., Streptococcus spp., Campylobacter spp., Mycobacterium spp., Helicobacter pylori, Staphylococcus spp., Staphylococcus aureus, Methicillin- sensitive Staphylococcus aureus (MSSA), Methicillin-resistant Staphylococcus aureus (MRSA), Pseudomonas aeruginosa and Escherichia coli, whereby the term "spp.”, as used herein, refers to a species.
  • the composition is used to treat or prevent an infection caused by, but is not limited to MRSA 1, MRSA 2, MRSA 3, MRSA 7, Mu50, WIS and MSSA 2590.
  • the described composition is used to treat a fungal infection.
  • the fungus may be, but is not limited to Candida albicans, Aspergillus brasiliensis and Aspergillus niger.
  • the present disclosure refers to a method of treating or preventing an infection in a patient, or for decolonizing an orifice of a patient, wherein the method comprised administering to the patient a therapeutically effective amount of the compositions as defined herein.
  • a composition as defined herein is used for treating or preventing an infection.
  • the use of a composition in the manufacture of a medicament for treating an infection is disclosed.
  • a method is disclosed wherein the composition is administered nasally via nasal irrigation or nasal fuming.
  • the present disclosure also refers to use of the composition as defined herein in the manufacture of a medicament for preventing or treating an infection, or for decolonizing an orifice of a patient.
  • the term “decolonization” refers to the local transient or permanent reduction of the amount of microorganisms in a patient. This decolonization may take place in, but is not limited to, a single, particular location in or on the body of the patient. Decolonization may take place in various locations in or on the body of the patient simultaneously or sequentially.
  • the term “orifice” refers to the entrance or outlet of any body cavity. Example of an orifice may be, but is not limited to, nose, ears, mouth, nostrils and throat.
  • the compositions, as described herein are used to treat or prevent an infection in an orifice, whereby the orifice may be, but is not limited to ear, nose, nasal cavity, sinus, nasal passageway, Eustachian tube and throat.
  • compositions described herein may be administered orally, topically, nasally, endosinually, intrasinally, transdermally, locally or combinations thereof.
  • nasal administration are, but not limited to, nasal irrigation, nasal fuming, nasal sprays and the like.
  • the compositions of the invention can also be administered by the nasal route.
  • the compositions may comprise a compound of the invention in a liquid carrier; such compositions may be administered for example in the form of a spray or as drops.
  • the liquid carrier may be water, which may contain further components to provide the desired isotonicity and viscosity of the composition.
  • the composition may also contain additional excipients such as preservatives, surface active agents and the like.
  • compositions may be contained in a nasal applicator that enables the composition to be administered as drugs or as a spray.
  • the composition should also include a propellant.
  • the composition may also be in unit dosage form, e.g. as tablets or capsules. In such form, the composition is sub-divided in unit dose containing appropriate quantities of the active ingredient; the unit dosage forms can be packaged composition, for example packeted powders, vials, ampoules, pre-filled syringes or sachets containing liquids.
  • the unit dosage form can be, for example, a capsule or tablet itself, or it can be the appropriate number of any such compositions in package form.
  • the present disclosure refers to the composition as disclosed herein, which may be provided in the form of, but not limited to dry powder, aqueous solution, non-aqueous solution, slurry, aerosols, nasal spray, nebulizer, inhalers, gels, creams, ointments, pastes, salves, suspensions, sterile solids, crystalline solids, amorphous solids, solids for reconstitution or combinations thereof.
  • the present disclosure refers to the composition as a nasal spray.
  • the composition is provided as a gel.
  • the composition is provided as a cream.
  • Nasal sprays are in liquid form such as an aqueous solution or suspension, an oil solution or suspension, or an emulsion, depending on the properties of the composition components.
  • Optional ingredients ensure minimal irritation, proper spray composition, and adequate delivery.
  • Buffers such as citrate, phosphate, and glycine adjust the pH of the nasal spray to prevent irritation to the nose.
  • Moisturizing agents such as propylene glycol and glycerine are also useful in the nasal spray.
  • polyphosphoesters such as polyethylene glycol, high molecular weight polylactic acid, microsphere encapsulations such as polyvinylpyrrolidone, hydroxypropyl cellulose, chitosan, and polystyrene sulfonate enhance the retention time of the composition.
  • microsphere encapsulations such as polyvinylpyrrolidone, hydroxypropyl cellulose, chitosan, and polystyrene sulfonate enhance the retention time of the composition.
  • each administration may comprise one or a plurality of applications or sprays of the claimed compositions, delivered to the nasal mucosa of the patient through one or both nostrils, the number of applications or sprays being dependent upon the concentration of the antimicrobial agent in the composition, the quantity of the composition delivered per spray, and the desired dose per administration as readily determined by one skilled in the art.
  • the composition can be dispensed from a spray bottle including a pump (e.g., a manually actuated pump) capable of delivering a metered spray of the composition of predetermined volume (typically about 0.1 mL).
  • a daily dose of 1% w/v of mupirocin in the composition may be administered in a single administration comprising one or more applications or metered sprays containing a total of 1% w/v of mupirocin (e.g., for 20 ⁇ g mupirocin, a single administration comprising two applications or metered sprays, one in each nostril and each containing 10 ⁇ g of mupirocin) or in multiple administrations (e.g., four administrations at six hour intervals, each administration comprising one or more applications or metered sprays, in one or both nostrils, each administration containing a total of 5 ⁇ g of mupirocin).
  • the antimicrobial composition When intended for use as an aerosol, the antimicrobial composition will be stored in and dispensed from a sealed container equipped with a metering valve and pump capable of being actuated to deliver or emit an aerosol (e.g., mist or spray) of the composition of predetermined volume into the patient's nostril and having a suitable droplet size distribution as known to those skilled in the art.
  • an aerosol e.g., mist or spray
  • the size of the droplets are large enough to prevent them from passing directly through the nasal passages and into the lungs, but small enough that they do not coalesce into large drops which either run out of the nose or down into the throat.
  • Suitable containers and metering valves for dispensing the antimicrobial composition according to the methods of the invention are available commercially and are known to those of skill in the art.
  • the container and valve system used to deliver the antimicrobial composition may incorporate any of the conventional aerosol formation techniques.
  • a low boiling liquid hydrocarbon such as, for example, butane, isobutane, propane, and other low boiling hydrocarbons in either pure or mixed forms
  • halohydrocarbon fluorocarbons (such as, for example, FC- 152A), chlorofluorocarbons (such as Freon or Freon like fluorocarbons, such as, for example, CFC-11, CFC-12 and CFC-114), and hydrofluorocarbons, also referred to as hydrofluoroalkanes (such as, for example, HFA-134a and HFA-227) are vaporized to exert a pressure and force the composition through the metering valve.
  • the antimicrobial composition can be stored for administration in a container or bottle including a pump and metering valve adapted for delivery of a metered spray of the composition and designed to inhibit or prevent degradation or spoilage of and bacterial growth in the composition contained therein.
  • nasal nebulisers are operated by pushing fumes through both nostrils. This mechanism results in a substantial amount of fumes being forced down into the lungs or lower respiratory tract, rather than to the inner nasal cavity.
  • the nasal irrigation has also been administered through one nostril at a time alternating between nostrils as a common clinical practice.
  • the nasal spray, irrigation or nebulised solution of antimicrobial compositions with or without sodium chloride (NaCl) may also be administered before an operation for nasal decolonization.
  • NaCl sodium chloride
  • Use of a saline nasal spray and irrigation several times per day has been shown in the art to help prevent scab formation in the nose.
  • composition described herein may also be used in form of a gel or a cream.
  • providing the composition as a gel or a cream allows for efficient, sterile and fuss-free application, for example, when the infection is present in the nose, the nostrils or within the nasal passageways of a patient.
  • the same also applies for applications for nasal decolonization in a patient, which may also utilize the compositions described herein, formulated or provided as gels or creams.
  • the composition comprises octenidine and cyclodextrin without a salt.
  • the composition comprises octenidine and cyclodextrin without a salt, wherein the composition is provided as a gel.
  • the composition comprises octenidine and cyclodextrin without a salt, wherein the composition is provided as a cream.
  • compositions of the antimicrobial agent with and without hypertonic saline increases the scope of application of the compositions.
  • alternative approaches to the standard of care in, for example, pre- and post-surgical nasal treatments by using, for example, spray or irrigation as a douche or a nebuliser to deliver fumes of antiseptic into the nasal cavity.
  • Methods, such as a douche or a nebulizer may be used to deliver fumes of antiseptic into the nasal cavity, for example during nose and sinus irrigations performed after functional sinus surgery.
  • the composition described herein may also have further secondary uses, such as but not limited to surgical sutures and for inhalation.
  • Non-medical uses such as, but not limited to, mouth wash, toothpaste, face wash, soaps, hand wash, surface disinfectants and cleaning products.
  • gel or cream formulations of octenidine dihydrochloride with or without sodium chloride can be used for antimicrobial applications.
  • composition described herein may be used in conjunction with other forms of treatment, or as adjunctive treatment.
  • the described composition may thus be administered before, after, during or together with a further treatment.
  • adjunctive treatment refers to another second treatment used together with the primary treatment, wherein the purpose of the secondary treatment is to assist the primary treatment.
  • each compound can be ground separately, after which they can be mixed together under constant agitation and heat to form the compositions as described herein.
  • the method may comprise mixing a ground cyclodextrin, as defined herein, and a ground antimicrobial agent, as defined herein, and constantly agitating and heating the resulting mixture. Heating and agitating can be done using means known in the art, e.g. in a thermostatically controlled shaker oven or water bath.
  • ground refers to the mechanical process of reducing the particle size of compounds or compositions, thereby producing a powdered form of the compound or composition. This grinding may be performed using methods known in the art, but generally encompass methods requiring that the compound be rubbed against a rough surface, whereby the roughness of the surface dictates how fine or coarse the resulting powder will be.
  • agitation may be replaced with “stirring” and refers to the constant moving of a mixture or a compound. "Constant” agitation can be applied to ensure the homogeneity of the resulting mixture, or to prevent the mixture from adhering to the sides of the mixing vessel during reaction.
  • the term "heat” refers to subject the compositions to a temperature that is higher than the ambient room temperature. Without being bound by theory, the inventors found that heating the composition described herein whilst constantly agitating it resulted in the antimicrobial agent being taken up into the pores of cyclodextrin, thus resulting in the complexation of the antimicrobial agent within cyclodextrin. This is also shown to be the case for antimicrobial agents that are known to be difficult to dissolve in, for example aqueous solutions. In order to be able to use this process, individual components of the compositions are subjected to heat.
  • these components are cyclodextrin, the antimicrobial agent or both cyclodextrin and the antimicrobial agent.
  • the temperature used in order to enable the complexation of the antimicrobial agent and cyclodextrin depends on the melting temperature of the antimicrobial agent. When heated to above this temperature, the antimicrobial agent becomes liquid and flows into the pores of cyclodextrin, thus enabling the complexation of the antimicrobial agent within cyclodextrin. However, it is to be noted that the heating temperature may not exceed the melting point of cyclodextrin, as this may inhibit the complexation of the antimicrobial agent with cyclodextrin.
  • the melting point of 2-hydroxypropyl-P-cyclodextrin is 278°C, and its degradation point is at around 300°C. Therefore, without being bound by theory, it would not be practical to melt cyclodextrin to form the complexation as there is a risk of degradation due to the proximity of its melting point to its degradation point. Economically, it is cheaper to use lower temperature to facilitate complexation of the compositions than higher temperature, as the lower temperature would require less energy. Thus, it will be more practical to melt the compound with low melting point to facilitate the complexation of the antimicrobial agent with cyclodextrin rather than to melt cyclodextrin for the complexation with the antimicrobial agent.
  • the heat applied in the method as described herein is the melting temperature of the antimicrobial agent.
  • the heat of applied may be between about 40°C to about 100°C, between about 50°C to about 200°C, between about 55°C to about 90°C, between about 60°C to about 80°C, between about 80°C to about 120°C, between about 70°C to about 90°C, between about 75°C to about 110°C, between about 95°C to about 145°C, between about 150°C to about 180°C, about 40°C, about 50°C, about 55°C, about 59°C, about 60°C, about 61°C, about 65°C, about 70°C, about 75°C, about 79°C, about 80°C, about 81°C, about 84°C, about 85°C, about 90°C, about 100°C, about 120°C, about 130°C, about 150°C, about 165°C, about 170°C, about 180°C or about 200°C.
  • the agitation time may be between about 0.5 hours to about 48 hours, between about 1 hour to about 48 hours, between about 1 hour to about 5 hours, between about 1 hour to about 10 hours, between about 5 hour to about 15 hours, between about 15 hour to about 20 hours, between about 18 hour to about 24 hours, between about 24 hour to about 36 hours, between about 36 hour to about 48 hours, between about 1 hour to about 1.5 hours, between about 1.5 hours to about 2 hours, between about 1.25 hours to about 2.25 hours, between about 2.5 hours to about 3 hours, about 1 hour, about 1.25 hours, about 1.75 hours, about 2.25 hours, about 2.75 hours, about 3 hours, about 4 hours, about 5 hours, about 10 hours, about 12 hours, about 20 hours, about 24 hours, about 28 hours, about 36 hours, about 40 hours, or about 48 hours.
  • the composition is agitated for between about 1 hour to about 24 hours.
  • the composition is agitated for 1 hour.
  • the composition is agitated for 2 hours.
  • the state of the compositions or mixtures may not only be ground, as previously described.
  • the state of the compositions may independently be dry or slurry, depending on the required downstream application.
  • slurry refers to a semi-liquid or fluid mixture, typically comprising fine particles of insoluble matter or matter with low solubility in a liquid solvent.
  • the solvent required for the formation of a slurry would vary depending on the components to be used and the downstream application. For example, in this method, water may be used as a solvent. In the case of water as a solvent, the term “slurry” would refer to a watery mixture. Another factor that may influence the formation of a slurry is the amount of solvent required.
  • the state of the mixture is dry or slurry. Therefore, the slurry formed by the method, as described herein, is a paste formed by adding a small amount of solvent to the mixed powder of the antimicrobial agent and cyclodextrin. This mixture is then agitated and/or triturated to form a slurry.
  • the term "triturate" refers to the process of grinding a compound or a mixture or a composition to a fine powder.
  • the methods described herein utilise the low melting temperatures of the antimicrobial compositions in order to enable complexation with cyclodextrin and enable the solubilisation of antimicrobial agents that otherwise may not have been possible.
  • octenidine will precipitate when mixed with sodium chloride in solution.
  • the methods described herein enable the mixing of compounds, for example, the antimicrobial agent and cyclodextrin, before the addition of sodium chloride resulted in a water-soluble composition without precipitation of any of the components.
  • the method disclosed herein may further comprise mixing the ground cyclodextrin with solvent to form a slurry; adding the ground antimicrobial agent to the slurry; subjecting the slurry to constant agitation as defined herein; grinding the resulting composition and mixing the resulting compositing with a salt as defined herein.
  • the disclosure refers to the method as described herein, wherein the antimicrobial agent is octenidine.
  • the term "about”, in the context of concentrations of components of the formulations, typically means + 5% of the stated value, more typically + 4% of the stated value, more typically + 3% of the stated value, more typically, + 2% of the stated value, even more typically + 1% of the stated value, and even more typically + 0.5% of the stated value.
  • range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the disclosed ranges. Accordingly, the description of a range should be considered to have specifically disclosed all the possible sub-ranges as well as individual numerical values within that range. For example, description of a range such as from 1 to 6 should be considered to have specifically disclosed sub-ranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6 etc., as well as individual numbers within that range, for example, 1, 2, 3, 4, 5, and 6. This applies regardless of the breadth of the range.
  • Octenidine dihydrochloride is formulated with 2-hydroxypropyl-P- cyclodextrin (HPpCD) using a slurry approach as described herein. This formulation can also be supplemented with 2.8% sodium chloride (NaCl). Stored as a dry powder, it can be reconstituted into an aqueous solution to be administered as irrigation or through a nebulizer for nasal and sinus cleansing and decolonization. The composition can be stored as a ready- to-use solution. The details on the methodology of preparing the composition comprising antimicrobial octenidine dihydrochloride are discusses herein.
  • compositions disclosed herein involve the use of cyclodextrins and its derivatives to enhance the solubility of mupirocin calcium (MPC) in water. 2.8% sodium chloride (NaCl) can also be added to the formulation. 2-hydroxypropyl-P-cyclodextrin (HPpCD) is chosen as it has low toxicity as compared to other cyclodextrins.
  • MPC mupirocin calcium
  • HPpCD 2-hydroxypropyl-P-cyclodextrin
  • HPpCD 2-hydroxypropyl-P-cyclodextrin
  • the formulation of MPC with and without hypertonic saline increases the scope of potential application of the formulation.
  • a dry powder containing triclosan (TCS) and 2-hydroxypropyl-P-cyclodextrin (HPPCD) is formulated using the respective aqueous and non-aqueous approach.
  • the dried powder is reconstituted into aqueous solution and administered to patients as nasal nebulization with any known nebulizer or simply as nasal douche by irrigation.
  • the composition can also be supplied as solution in ampoules for reconstitution by dilution for the same nasal nebulization or nasal irrigation.
  • a suitable method to administering the composition by nebulisation is provided.
  • the composition can be stored as a dry powder, ready-to-use solution, or concentrated composition. The process for preparation of triclosan nebulising solution is also described herein.
  • octenidine-cyclodextrin (OCT-CD) inclusion complexes For the preparation of, for example, octenidine-cyclodextrin (OCT-CD) inclusion complexes, a 'slurry complexation-dry heat' method is used.
  • 2-hydroxypropyl-P- cyclodextrin (HPpCD; Cavasol W7 HP Pharma, Wacker Chemicals) and the antimicrobial agent were weighed out and ground in a glass mortar.
  • water and 0.1N sodium hydroxide (NaOH) were added per milligram of ground mixture (for example, 10ml of lmg/ml octenidine in water). This allows the dissolution of cyclodextrin and the formation of a viscous slurry with constant stirring.
  • the slurry was then dried in a thermostatically controlled shaker oven at 80 + 1°C for 1 hour, in order to remove any access water from the slurry and thereby forming a powder.
  • the temperature of the controlled shaker oven was adjusted according to the melting point of the antimicrobial agent.
  • the resulting powder was then ground to ensure uniformity.
  • 2.8% w/v sodium chloride (NaCl) was weighed and ground together with the dried slurry powder.
  • HPLC High Performance Liquid Chromatography
  • Chromatographic separation was achieved on Mightysil RP-18 GP 150 mm X 4.6 mm, particle size 5 ⁇ (Kanto Chemical Co.) analytical column with an isocratic mobile phase of 70% acetonitrile and 30% water, whereby the composition of the mobile phase is adjusted according to the sample in question. HPLC grade acetonitrile was used. All parameters for chromatographic separation were also adjusted according to the physic- chemical properties of the antimicrobial agent in question, according to methods known in the art. For example, a flow rate of 1.2 ml/min was used and the UV detection wavelength was set at 243 nm to monitor analyte peak elution. Flow rate and UV detection wave length are also adjusted according to the sample to be analysed. The oven temperature was maintained at 50°C.
  • the viscosities of the compositions in solution were evaluated using the parallel plate method.
  • the rheometer used was the ARES-G2 Rheometer, TA Instruments, USA.
  • the plate temperature was set at 25 °C.
  • a total of 100 data points was measured with a shear rate from 0.5 s "1 to 60 s "1 , for a total time of 10 minutes per run.
  • the viscosities of MilliQ water and the complexes containing 0.25% to 5% CMC were obtained and plotted in a graph of shear stress (Pa) against shear rate (s _1 ).
  • Undissolved antimicrobial agent was removed by filtration using 0.22 ⁇ Millex® GP Filter Units. The resulting solution was then diluted appropriately and quantified using the HPLC assay. The saturation of the antimicrobial agent in water without sodium chloride and antimicrobial agent in water with 2.8% sodium chloride (NaCl) was also evaluated as a control. After analysing the results of the saturation study, 2-hydroxypropyl-P- cyclodextrin (HPpCD) at a concentration of 50 mM was chosen to formulate compositions of an antimicrobial agent in the presence of 2.8% NaCl.
  • HPpCD 2-hydroxypropyl-P- cyclodextrin
  • 2-hydroxypropyl-P- cyclodextrin had greater solubility than a-cyclodextrin in water, with the maximum solubility at 300 mM and 100 mM respectively. Furthermore, at 50 mM, it was sufficient to dissolve more than 10 times a target concentration of antimicrobial agent (set at 1 mg/ml) in the presence of hypertonic saline. At this 2-hydroxypropyl-P-cyclodextrin concentration, the antimicrobial composition could be solubilised quickly and the slurry could be manipulated more easily due to its lower viscosity.
  • MIC Minimum Inhibitory Concentration
  • a series of 15 mL glass tubes were filled with double- strength (D/S) nutrient broth (Acumedia® Nutrient Broth 7146, Lot: 106490B), placed in a test-tube rack and labelled accordingly.
  • the composition to be analysed is dissolved in MilliQ water and filtered through 0.22 ⁇ Millex® GP Filter Units. Varying volumes of the dissolved composition were added to each glass tube giving a range of concentrations for the determination of the minimum inhibitory concentration (MIC). Sterile water was then added to each glass tube. The tubes were then mixed thoroughly by rotating between the palms of the hands.
  • 0.2 ml of a diluted 24-hour broth culture of Staphylococcus aureus (MicroBiologics® S. aureus; Lot No.: 827963; RFF 0827S; ATCC 6538P) standardised at 0.5 McFarland Standard, was next added to each tube.
  • a positive control containing only the cultured organism, sterile water and double-strength (D/S) nutrient broth was also prepared. The tubes were then mixed thoroughly again by rotating between the palms of the hands, and incubated at 37°C for 72 hours.
  • the tubes were observed for turbidity, indicative of microbiological growth at 72 hours and quantified by comparing against McFarland Equivalence Standards (Batch No.: 1313947; TM4000T10).
  • the minimum inhibitory concentration was recorded as the lowest antimicrobial concentration where no bacteria growth is observed after 72 hours incubation. All apparatus (i.e. glass tubes, measuring cylinders, pipette tips, stock bottles, and volumetric flasks), nutrient broth solution and sterile water were placed through a moist heat sterilisation process (autoclave) at 121°C for 15 minutes before they were used.
  • Each set of tubes included a positive (bacteria without the addition of antimicrobial formulation) and negative control (broth only). The experiments were performed in duplicates.
  • MIC Minimum Inhibitory Concentration
  • the minimum inhibitory concentration (MIC) of a composition is determined against 7 Staphylococcus aureus strains: MRSA 1, MRSA 2, MRSA 3, MRSA 7, Mu50, WIS and MSSA 2590 using the Minimum Inhibitory Concentration (MIC) Broth Microdilution Assay.
  • the assay was conducted in accordance to the guidelines set out in 'Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria that Grow Aerobically - Eighth Edition' (Clinical and Laboratory Standards Institute, 2012). All Staphylococcus aureus strains were kindly provided by Jeanette Teo from National University Hospital.
  • MRSA 1, MRSA 2, MRSA 3 and MRSA 7 are clinically isolated MRSA strains.
  • MRSA strains Mu50, WIS and MSSA strain 2590 were obtained from ATCC. Out of the clinically-isolated MRSA strains, MRSA 1 and MRSA 2 are resistant to mupirocin. They are characterised by the presence of the mupA gene.
  • the drug compositions were dissolved in MilliQ water and filtered through 0.22 ⁇ Millex® GP Filter Units. It was further diluted in Mueller Hinton (MH) broth (Ref: 275730, Lot: 0224378) to appropriate concentrations for the determination of the MIC.
  • MH Mueller Hinton
  • Sterile 96-well round-bottom microtiter plates (Nunc, Roskilde, Denmark) were used. 24- hour broth culture of S. aureus was diluted with MH broth (and standardised at between 1 X 105 - 1 X 106 CFU/ml. This was determined by the plating of the bacteria on LB agar at the time of inoculation. Each row of the microplate was loaded with a different S. aureus strain.
  • test inoculum 50 ⁇ of test inoculum was loaded into each well. 50 ⁇ of drug solution was then added into first column and serial two-fold dilution is performed across the columns. The final volume of the well was 50 ⁇ . The 96-well microtiter plates were then covered with a plate cover and incubated overnight at 37°C. The minimum inhibitory concentration was recorded as the lowest antimicrobial concentration where no bacteria growth is observed after overnight incubation. The experiments were performed in duplicates.
  • compositions according to the invention were weighed and dissolved in 10 ml MilliQ water. Following that, the samples were filtered using 0.22 ⁇ Millex® GP Filter Units. The stability of the samples was evaluated for 8 weeks under two conditions:
  • the time-dependent bactericidal kill profile of the compositions was determined at full-strength and half-strength against Staphylococcus aureus strains: MRS A 1, MRS A 2, MRSA7 and Mu50.
  • the compositions were dissolved directly in MH broth in a 15ml falcon tube.
  • Half-strength formulations were prepared by performing a two-fold dilution of the full- strength formulations.
  • test inoculums were conducted using suitable mediums (Soybean-Casein Digest Broth and Sabouraud Dextrose Broth).
  • the test microorganisms include: Escherichia coli (ATCC 8739), Pseudomonas aeruginosa (ATCC 9027), Staphylococcus aureus (ATCC 6538), Candida albicans (ATCC 10231) and Aspergillus brasiliensis (ATCC 16404).
  • the test inoculums were incubated for microbial recovery.
  • the incubated test inoculums were standardised such that the final concentration of the test preparations after inoculation is between lxlO 5 and lxlO 6 CFU/ml of the product.
  • the initial concentration of viable test microorganisms in each test preparation is determined by the plate-count method.
  • Each container was sampled and enumerated using the plate-count procedure at applicable test intervals: Day 1, Day 7, Day 14 and Day 28 of incubation.
  • the logio values of the concentration of each microorganism at the applicable test intervals were calculated and reported to investigate the log reductions.
  • the log reduction is defined as the difference between the loglO unit value of the starting concentration (in CFU/ml) in the suspension and the log 10 unit value of the surviving concentration (in CFU/ml) at that time point.
  • Bacteria Not less than 1.0 log reduction from the initial calculated count at 7 days, and not less than 3.0 log reduction from the initial count at 14 days, and no increase from the 14 days' count at 28 days.
  • the main components of the formulation were 2-hydroxypropyl-P-cyclodextrin (HPPCD), sodium carboxymethyl cellulose (CMC), crystalline sodium chloride (NaCl), the poorly water-soluble phenolic antiseptic, triclosan (TCS), mupirocin calcium (MPC) and octenidine dihydrochloride (OCT).
  • HPPCD 2-hydroxypropyl-P-cyclodextrin
  • CMC carboxymethyl cellulose
  • NaCl crystalline sodium chloride
  • TCS poorly water-soluble phenolic antiseptic
  • MPC mupirocin calcium
  • OCT octenidine dihydrochloride
  • Example 1 Non-Aqueous Approach (Dry physical mixing) to mupirocin compositions
  • Amorphous mupirocin characterised by a melting point of about 77°C to about 89° may be prepared by the same dry heat approach to melt mupirocin and triturate with 2-hydroxypropyl-P-cyclodextrin or other cyclodextrins and cyclodextrin derivative.
  • the mixed dried powder may be reconstituted into solution for nasal nebulization or irrigation.
  • the composition maybe modified by adding CMC to enhance the solubility and/or cohesive effects to the nasal membrane.
  • Sodium chloride may also be added to the formulation for the additive/synergistic antimicrobial effects.
  • mupirocin-cyclodextrin inclusion complexes For the preparation of mupirocin-cyclodextrin inclusion complexes, a 'dry physical mixing' method is used. In this method, 100 mM of 2-hyrdroxypropyl-P-cyclodextrin (HPPCD) and 2% w/v mupirocin calcium (MPC) were weighed out and ground in a glass mortar. Following that, the ground physical mixture was subject to constant agitation in glass vials at 275 rpm in a thermostatically controlled shaker oven at 85 + 1°C for 2 hours. This allowed mupirocin calcium to melt into a liquid (melting point of 77 - 78°C), which facilitated the incorporation into the cyclodextrin cavities.
  • HPPCD 2-hyrdroxypropyl-P-cyclodextrin
  • MPC 2% w/v mupirocin calcium
  • MPC-HPpCD was obtained by dissolving the dry formulation of MPC-HPpCD which consists of 20 mg/ml mupirocin calcium and 100 mM 2-hydroxypropyl- ⁇ -cyclodextrin.
  • MPC-HPpCD-NaCl was obtained by dissolving the dry formulation of MPC-HPpCD-NaCl which consists of 20 mg/ml mupirocin calcium (MPC), 100 mM 2-hydroxypropyl-P-cyclodextrin (HPpCD) and 2.8% w/v sodium chloride (NaCl). Formulations with 0.25% w/v carboxymethyl cellulose sodium (CMC) were also evaluated.
  • MPC-HPpCD-CMC was obtained by dissolving the dry formulation of MPC-HPpCD-CMC which consists of 20 mg/ml mupirocin calcium (MPC), 100 mM 2-hydroxypropyl-P- cyclodextrin (HPpCD) and 0.25% w/v carboy methyl cellulose (CMC).
  • MPC-HPpCD- CMC-NaCl was obtained by dissolving the dry formulation of MPC-HPpCD-CMC-NaCl which consists of 20 mg/ml mupirocin calcium (MPC), 100 mM 2-hydroxypropyl-P- cyclodextrin (HPpCD), 2.8% w/v sodium chloride (NaCl) and 0.25% w/v carboxymethyl cellulose (CMC).
  • MPC-HPpCD-CMC-NaCl which consists of 20 mg/ml mupirocin calcium (MPC), 100 mM 2-hydroxypropyl-P- cyclodextrin (HPpCD), 2.8% w/v sodium chloride (NaCl) and 0.25% w/v carboxymethyl cellulose (CMC).
  • Example 2 Preparation of 2.0% w/v mupirocin calcium (MPC) in 100 mM 2 -hydroxy propy l- ⁇ -cyclodextrin ( ⁇ )
  • mupirocin-cyclodextrin (MPC-CD) inclusion complexes For the preparation of mupirocin-cyclodextrin (MPC-CD) inclusion complexes, the 'dry physical mixing' method is used. In this method, 1400 mg of 2-hydroxypropyl-P- cyclodextrin (HPpCD) and 200 mg mupirocin (MPC) were weighed out and ground in a glass mortar. Following that, the ground physical mixture was subject to constant agitation in glass vials at 275 rpm in a thermostatically controlled shaker oven at 85 + 1°C for 2 hours. This allowed mupirocin to melt into a liquid (melting point of 77 - 78°C), which facilitated the incorporation into the cyclodextrin cavities.
  • HPpCD 2-hydroxypropyl-P- cyclodextrin
  • MPC mupirocin
  • Example 3 Preparation of 2.0% w/v mupirocin chloride (MPC) in 100 mM 2-hydroxypropyl- ⁇ -cyclodextrin ( ⁇ ) with 2.8% w/v sodium chloride (NaCl) [0133]
  • MPC-CD mupirocin-cyclodextrin
  • HPpCD 2-hydroxypropyl-P- cyclodextrin
  • MPC 280 mg sodium chloride
  • NaCl sodium chloride
  • the ground physical mixture was subject to constant agitation in glass vials at 275 rpm in a thermostatically controlled shaker oven at 85 + 1°C for 2 hours. This allowed mupirocin chloride to melt into a liquid (melting point of 77 - 78 °C), which facilitated the incorporation into the cyclodextrin cavities.
  • the heated and agitated mixture was then allowed to cool to room temperature (25°C). After cooling, the mixture was ground again for 10 minutes to ensure uniformity of mixture and facilitate rapid reconstitution later.
  • Example 4 Preparation of 2.0% w/v mupirocin calcium (MPC) in 100 mM HPfCD with 2.8% w/v sodium chloride (NaCl) and 0.25% w/v carboxymethylcellulose sodium (CMC)
  • mupirocin-cyclodextrin (MPC-CD) inclusion complexes For the preparation of mupirocin-cyclodextrin (MPC-CD) inclusion complexes, the 'dry physical mixing' method is used. In this method, 1400 mg of 2-hydroxypropyl-P- cyclodextrin (HPpCD), 200 mg mupirocin calcium (MPC), 280 mg sodium chloride (NaCl) and 25 mg carboxymethyl cellulose (CMC) were weighed out and ground in a glass mortar. Following that, the ground physical mixture was subject to constant agitation in glass vials at 275 rpm in a thermostatically controlled shaker oven at 85 + 1°C for 2 hours.
  • HPpCD 2-hydroxypropyl-P- cyclodextrin
  • MPC 2-hydroxypropyl-P- cyclodextrin calcium
  • NaCl sodium chloride
  • CMC carboxymethyl cellulose
  • the method is performed as previously outlined.
  • the retention time of mupirocin calcium (MPC) was 1.2 min with peak width of 0.4 min.
  • Table 1 High Performance Liquid Chromatography (HPLC) Assay Parameters for Mupirocin.
  • mupirocin was forcibly degraded by acid (IN hydrochloric acid) and base (IN sodium hydroxide) at room temperature and at 40°C for an hour. Both drugs were also placed under UV for 24 hours. Both drugs were dissolved in 50% acetonitrile and 50% water in screw cap glass tubes and for each degradation condition, triplicates were prepared. The samples were neutralised with an equal amount of IN sodium hydroxide and IN hydrochloric acid respectively. The samples were diluted in mobile phase before analysis. No degradation product peak was obtained for mupirocin under UV light for 24 hours. Degradation products were observed under acid and base degradation conditions at retention times of 1.2 min, 1.35 min and 5.0 min.
  • the MPC-HPpCD-NaCl composition was weighed and dissolved in 10 ml MilliQ water. Following that, it was filtered using 0.22 ⁇ Millex® GP Filter Units. The stability of the formulation was evaluated for 8 weeks under two conditions:
  • Example 8 Mupirocin Saturation in 2-hydroxypropyl-fi-cyclodextrin ( ⁇ ) and a- cyclodextrin (a-CD)
  • HPpCD 2-hydroxypropyl-P-cyclodextrin
  • a series of 15 mL glass tubes were filled with double- strength (D/S) nutrient broth (Acumedia® Nutrient Broth 7146, Lot: 106490B), placed in a test-tube rack and labelled accordingly.
  • D/S double- strength
  • Four different formulations (MPC-HPpCD, MPC-HPpCD-NaCl, MPC-HPpCD- CMC, and MPC-HPpCD-CMC-NaCl, prepared by dry physical mixing) were dissolved in MilliQ water and filtered through 0.22 ⁇ Millex® GP Filter Units. Varying volumes of these formulations were added to each glass tube giving a range of concentrations for the determination of the minimum inhibitory concentration (MIC). Sterile water was then added to each glass tube.
  • MIC minimum inhibitory concentration
  • 0.2 ml of a diluted 24-hour broth culture of Staphylococcus aureus (MicroBiologics® S. aureus; Lot No.: 827963; RFF 0827S; ATCC 6538P) standardised at 0.5 McFarland Standard, was next added to each tube.
  • a positive control containing only the cultured organism, sterile water and double-strength (D/S) nutrient broth was also prepared. The tubes were then mixed thoroughly again by rotating between the palms of the hands, and incubated at 37°C for 72 hours.
  • the tubes were observed for turbidity, indicative of microbiological growth at 72 hours and quantified by comparing against McFarland Equivalence Standards (Batch No.: 1313947; TM4000T10).
  • the minimum inhibitory concentration was recorded as the lowest antimicrobial concentration where no bacteria growth is observed after 72 hours incubation.
  • MIC Minimum Inhibitory Concentration
  • MRSA 1, MRSA 2, MRSA 3 and MRSA 7 are clinically isolated MRSA strains.
  • MRSA strains Mu50, WIS and MSSA strain 2590 were obtained from ATCC.
  • MRSA 1 and MRSA 2 are resistant to mupirocin. They are characterised by the presence of the mupA gene.
  • the drug compositions were dissolved in MilliQ water and filtered through 0.22 ⁇ Millex® GP Filter Units. It was further diluted in Mueller Hinton (MH) broth (Ref: 275730, Lot: 0224378) to appropriate concentrations for the determination of the MIC.
  • MH Mueller Hinton
  • Sterile 96-well round-bottom microtiter plates (Nunc, Roskilde, Denmark) were used. 24-hour broth culture of S. aureus was diluted with MH broth (and standardised at between lxlO 5 - lxlO 6 CFU/ml. This was determined by the plating of the bacteria on LB agar at the time of inoculation. Each row of the microplate was loaded with a different S. aureus strain.
  • test inoculum 50 ⁇ of test inoculum was loaded into each well. 50 ⁇ of drug solution was then added into first column and serial two-fold dilution is performed across the columns. The final volume of the well was 50 ⁇ . The 96-well microtiter plates were then covered with a plate cover and incubated overnight at 37°C. The minimum inhibitory concentration was recorded as the lowest antimicrobial concentration where no bacteria growth is observed after overnight incubation. The experiments were performed in duplicates.
  • MIC Minimum Inhibitory Concentration
  • the minimum inhibitory concentration (MIC) of the MPC -based formulation was found to be between 0.1 to 0.2 ⁇ g/ml for Methicillin-resistant Staphylococcus aureus (MRSA) and Methicillin-sensitive Staphylococcus aureus (MSSA) bacteria strains. MIC increases from 48 to 96 ⁇ g/ml for MPC-resistant MRSA bacteria strains.
  • Example 11 Antibacterial effectiveness of mupirocin compositions
  • test inoculums were conducted using suitable mediums and in accordance to the method as previously described.
  • the test microorganisms include: Escherichia coli (ATCC 8739), Pseudomonas aeruginosa (ATCC 9027), Staphylococcus aureus (ATCC 6538), Candida albicans (ATCC 10231) and Aspergillus brasiliensis (ATCC 16404).
  • the test inoculums were incubated for microbial recovery.
  • the incubated test inoculums were standardised such that the final concentration of the test preparations after inoculation is between lxlO 5 and lxlO 6 CFU/ml of the product.
  • the initial concentration of viable test microorganisms in each test preparation is determined by the plate-count method.
  • Example 12 Time-dependent bacterial activity of mupirocin compositions
  • the time-dependent bactericidal kill profile of two mupirocin calcium compositions (MPC-HPpCD, MPC-HPpCD-NaCl) at full-strength and half-strength were determined against Staphylococcus aureus strains: MRS A 1, MRS A 2, MRSA7 and Mu50.
  • the compositions were dissolved directly in MH broth in a 15ml falcon tube.
  • Half-strength formulations were prepared by performing a two-fold dilution of the full- strength formulations.
  • Example 13 Slurry approach to octenidine compositions
  • octenidine-cyclodextrin (OCT-CD) inclusion complexes For the preparation of octenidine-cyclodextrin (OCT-CD) inclusion complexes, a 'slurry complexation-dry heat' method is used. In this method, 50 mM of 2-hydroxypropyl-P- cyclodextrin (HPpCD; Cavasol W7 HP Pharma, Wacker Chemicals), 0.1% w/v octenidine dihydrochloride (Tokyo Chemical Industry) were weighed out and ground in a glass mortar. Following that, 200 ⁇ of water and 3.5 ⁇ of 0.1N sodium hydroxide (NaOH) were added per 710 mg of ground mixture (i.e. 10ml of lmg/ml octenidine in water).
  • HPpCD 2-hydroxypropyl-P- cyclodextrin
  • NaOH 0.1N sodium hydroxide
  • the slurry was then dried in a thermostatically controlled shaker oven at 80 + 1°C for 1 hour. The resulting powder was then ground to ensure uniformity. In the case of formulation with hypertonic saline, 2.8% w/v sodium chloride (NaCl) was weighed and ground together with the dried slurry powder.
  • NaCl sodium chloride
  • Octenidine dihydrochloride-HiO was obtained by dissolving 1 mg/ml of octenidine dihydrochloride in water.
  • Octenidine dihydrochloride-HPpCD solution was obtained by dissolving the dry formulation of OCT- HPPCD which consists of 1 mg/ml octenidine dihydrochloride and 50 mM 2-hydroxypropyl- ⁇ -cyclodextrin (HPpCD).
  • OCT-HPpCD-NaCl solution was obtained by dissolving the dry formulation of OCT-HPpCD-NaCl which consists of 1 mg/ml octenidine hydrochloride (OCT), 50 mM 2-hydroxypropyl-p-cyclodextrin (HPpCD) and 2.8% sodium chloride (NaCl).
  • OCT-HPpCD-NaCl consists of 1 mg/ml octenidine hydrochloride (OCT), 50 mM 2-hydroxypropyl-p-cyclodextrin (HPpCD) and 2.8% sodium chloride (NaCl).
  • Example 14 Preparation of 0.1% w/v octenidine in 50 mM 2-hydroxypropyl-fi-cyclodextnn 10 mg octenidine dihydrochloride (Tokyo Chemical Industry) and 700 mg 2-hydroxypropyl- ⁇ -cyclodextrin (HPpCD) (Cavasol W7 HP Pharma, Wacker Chemicals) was weighed out and ground in a glass mortar. Following that, 200 ⁇ 1 of water and 3.5 ⁇ of 0.1 N sodium hydroxide (NaOH) were added. This allowed the dissociation of cyclodextrin and viscous slurry was formed with constant stirring. The slurry was then dried in a thermostatically controlled shaker oven at 80 + 1°C for 1 hour.
  • Tokyo Chemical Industry 2-hydroxypropyl-fi-cyclodextnn 10 mg octenidine dihydrochloride (Tokyo Chemical Industry) and 700 mg 2-hydroxypropyl- ⁇ -cyclodextrin (HPpCD) (Cava
  • the resulting powder was then ground to ensure uniformity.
  • the dried powder can be readily reconstituted with 10 ml water to give 0.1% w/v octenidine dihydrochloride in 50 mM 2-hydroxypropyl-P-cyclodextrin.
  • Example 15 Preparation of 0.1% w/v octenidine in 50 mMa -cyclodextrin
  • Example 16 Preparation of 0.1% w/v octenidine in 50 mM 2-hydroxypropyl-fi-cyclodextrin ( ⁇ ) with 2.8% w/v sodium chloride (NaCl)
  • the dried powder can be readily reconstituted with 10 ml water to give 0.1% w/v octenidine dihydrochloride in 50 mM 2-hydroxypropyl-P-cyclodextrin (HPpCD) with 2.8% w/v sodium chloride (NaCl).
  • Example 17 Preparation of 0.1% w/v octenidine in 50 mM a-cyclodextrin with 2.8% w/v sodium chloride (NaCl)
  • the dried powder can be readily reconstituted with 10 ml water to give 0.1%) w/v octenidine dihydrochloride (OCT) in 50 mM ⁇ -cyclodextrin (a-CD) with 2.8% w/v sodium chloride (NaCl).
  • OCT octenidine dihydrochloride
  • a-CD ⁇ -cyclodextrin
  • NaCl sodium chloride
  • HPLC High Performance Liquid Chromatography
  • octenidine dihydrochloride was forcibly degraded by acid (IN hydrochloric acid, HCl) and base (IN sodium hydroxide, NaOH) at room temperature and at 40°C for an hour. Both drugs were also placed under UV for 24 hours. Both drugs were dissolved in 50% acetonitrile and 50% water in screw cap glass tubes and for each degradation condition, triplicates were prepared. The samples were neutralised with an equal amount of IN sodium hydroxide (NaOH) and IN hydrochloric acid (HCl) respectively. The samples were diluted in mobile phase before analysis. No degradation product peak was obtained for octenidine under these conditions.
  • the parent ion or Ql scan, conducted at the first quadrupole mass filter was performed to identify degradation in octenidine dihydrochloride. Degradation can be concluded if there is a reduction in parent ion intensity or if there were other prominent peaks other than the parent ion peak.
  • the stability of the formulation was evaluated after 8 weeks of storage. The dry powder OCT- HPpCD-NaCl formulation is observed to be stable for up to 8 weeks (the stability study is still on going to establish the long-term stability).
  • Example 21 Octenidine saturation in 2-hydroxypropyl-fi-cyclodextrin ( ⁇ ) and a- cyclodextrin (a-CD)
  • Undissolved octenidine dihydrochloride was removed by filtration using 0.22 ⁇ Millex® GP Filter Units. The resulting solution was then diluted appropriately and quantified using the HPLC assay. The saturation of octenidine dihydrochloride in water and octenidine in water with 2.8% sodium chloride (NaCl) was also evaluated as a control. After analysing the results of the saturation study, 2-hydroxypropyl-P- cyclodextrin (HPpCD) at a concentration of 50 mM was chosen to formulate octenidine dihydrochloride in the presence of 2.8% NaCl.
  • HPpCD 2-hydroxypropyl-P- cyclodextrin
  • Example 22 Minimum Inhibitory Concentration (MIC) of octenidine compositions- Macrodilution ( tube ) broth assay
  • 0.2 ml of a diluted 24-hour broth culture of Staphylococcus aureus (MicroBiologics® S. aureus; Lot No.: 827963; RFF 0827S; ATCC 6538P) standardised at 0.5 McFarland Standard, was next added to each tube.
  • a positive control containing only the cultured organism, sterile water and double-strength (D/S) nutrient broth was also prepared. The tubes were then mixed thoroughly again by rotating between the palms of the hands, and incubated at 37°C for 72 hours.
  • the tubes were observed for turbidity, indicative of microbiological growth at 72 hours and quantified by comparing against McFarland Equivalence Standards (Batch No.: 1313947; TM4000T10).
  • the minimum inhibitory concentration was recorded as the lowest antimicrobial concentration where no bacteria growth is observed after 72 hours incubation.
  • Example 23 Minimum Inhibitory Concentration (MIC) of octenidine compositions- Microdilution ( tube ) broth assay
  • MIC Minimum inhibitory concentration
  • OCT-HPpCD OCT-HPpCD-NaCl
  • MRSA 1, MRSA 2, MRSA 3, MRSA 7, Mu50, WIS and MSSA 2590 The minimum Inhibitory Concentration (MIC) Broth Microdilution Assay.
  • the assay was conducted in accordance to the guidelines set out in 'Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria that Grow Aerobically - Eighth Edition' (Clinical and Laboratory Standards Institute, 2012).
  • MRSA 1 and MRSA 2 are clinically isolated MRSA strains.
  • MRSA strains Mu50, WIS and MSSA strain 2590 were obtained from ATCC.
  • MRSA 1 and MRSA 2 are resistant to mupirocin. They are characterised by the presence of the mupA gene.
  • the drug formulations were dissolved in MiUiQ water and filtered through 0.22 ⁇ Millex® GP Filter Units. It was further diluted in MH broth to appropriate concentrations for the determination of the MIC. Sterile 96-well round-bottom microtiter plates (Nunc, Roskilde, Denmark) were used. 24-hour broth culture of S. aureus was diluted with MH broth (and standardised at between lxlO 5 - lxlO 6 CFU/ml. This was determined by the plating of the bacteria on Luria Broth (LB) Agar at the time of inoculation. Each row of the microplate was loaded with a different S. aureus strain. 50 ⁇ of test inoculum was loaded into each well.
  • LB Luria Broth
  • MIC Minimum Inhibitory Concentration
  • the minimum inhibitory concentration (MIC) of the octenidine dihydrochloride- based formulation was found to be about 1.6 Mg/ml for all Methicillin -resistant Staphylococcus aureus (MRSA) and Methicillin-sensitive Staphylococcus aureus (MSSA) bacteria strains tested. It was also found to have equal activity against mupirocin-resistant and mupirocin-responsive MRSA.
  • MRSA Methicillin -resistant Staphylococcus aureus
  • MSSA Methicillin-sensitive Staphylococcus aureus
  • Example 24 Time-dependent determination of bacterial activity of octenidine compositions
  • the time-dependent bactericidal kill profile of two compositions (OCT-HPpCD, OCT-HPpCD-NaCl) at full-strength and half-strength were determined against Staphylococcus aureus strains: MRSA 1, MRSA 2, MRSA7 and Mu50.
  • the formulations were dissolved directly in MH broth in a 15ml falcon tube.
  • Half-strength formulations were prepared by performing a two-fold dilution of the full-strength formulations. At time zero, 2.5 ⁇ of 24-hour broth culture of S.
  • aureus was added to 5 ml of each formulation such that the bacteria concentration is between 1x105 - 1x106 CFU/ml (colony forming units per ml).
  • MH Mueller-Hinton
  • Example 25 Antimicrobial effectiveness testing of octenidine compositions against bacteria and fungi
  • Example 26 Aqueous (slurry complexation) and non-aqueous (dry heat mixing) approach to triclosan compositions
  • the completed slurries were collected in 1.5 ml Eppendorf® tubes and placed in a freeze-dryer (0.05 mBar, -47°C) for 2 hours. The dried powders were obtained and each was dissolved in 10 mL of MilliQ water, which were filtered immediately through 0.22 ⁇ Millex® GP Filter Units.
  • Example 27 Dry physical mixture of HPfCD-CMC-TCS for nasal nebulisation
  • This triclosan-based solution composition can be used in a nebuliser to deliver fumes of antiseptic to the nasal cavities, providing decolonising effect.
  • 2-hydroxypropyl-P-cyclodextrin and carboxymethyl cellulose serve to improve the aqueous solubility of the formulation and modulate the release of triclosan in the nasal cavity when nebulised.
  • Example 28 Dry physical mixture of HP fCD-CMC-TCS-NaCl for nasal nebulisation
  • This triclosan-based solution composition can be used in a nebuliser to deliver fumes of antiseptic to the nasal cavities, providing decolonising effect.
  • 2-hydroxypropyl-P-cyclodextrin (FIPpCD) and carboxymethyl cellulose (CMC) serve to improve the aqueous solubility of the formulation and modulate the release of triclosan in the nasal cavity when nebulised.
  • Saline is added for its bacteriostatic and irrigative properties, and also enhances the aqueous solubility of the formulation.
  • Example 29 Lyophilised mixture of HPfCD-CMC-TCS for nasal nebulisation
  • This triclosan-based solution composition can be used in a nebuliser to deliver fumes of antiseptic to the nasal cavities, providing decolonising effect.
  • 2-hydroxypropyl-P- cyclodextrin (FfPpCD) and carboxymethyl cellulose serve to improve the aqueous solubility of the formulation and modulate the release of triclosan in the nasal cavity when nebulised.
  • Example 30 Lyophilised mixture ofHPfiCD-CMC-TCS-NaClfor nasal nebulisation.
  • HPPCD, TCS, NaCl and CMC were added in sequence in order to produce slurry for lyophilisation.
  • a small quantity of water (less than 500 ⁇ ,) was added to dissolve FfPpCD and stirred with constant agitation for about 15 minutes, until a clear solution was obtained.
  • TCS was then added to the aqueous CD and ground for another 30 minutes, producing a cloudy paste.
  • NaCl was added to the paste next, and further grinding incorporated the crystals into the slurry.
  • the addition of CMC produced a viscous and sticky paste, which was then collected in Eppendorf® tubes and placed in a freeze-dryer (0.05 mBar, -47°C) for 24 hours.
  • Example 31 High Performance Liquid Chromatography (HPLC) of triclosan compositions
  • HPLC High Performance Liquid Chromatography
  • Example 32 Stability Assay validation of triclosan compositions
  • triclosan was forcibly degraded by acid (IN hydrochloric acid, HC1) and base (IN sodium hydroxide, NaOH) at room temperature and at 40°C for an hour. Both acidic and basic samples were also placed under UV light for 24 hours. Both drugs were dissolved in 50% acetonitrile and 50% water in screw cap glass tubes and for each degradation condition, triplicates were prepared. The samples were neutralised with an equal amount of IN sodium hydroxide (NaOH) and IN hydrochloric acid (HC1) respectively. The samples were diluted in mobile phase before analysis. No degradation product peak was obtained for triclosan under these conditions.
  • Example 33 Triclosan powder composition stability assay
  • the stability of powder TCS-HPpCD-NaCl composition was evaluated with the QTRAP 5500 LC-MS/MS system.
  • the formulation was reconstituted by dissolving in MilliQ water and diluted to a concentration of 1 ⁇ g/ml with 50% methanol and 50% water.
  • the solution was directly injected into the LC-MS/MS system and the parent ion was scanned with in the negative scanning mode and a scan range of 150 to 300.
  • the triclosan parent ion peak was detected at m/z of 288.9.
  • the parent ion or Ql scan, conducted at the first quadrupole mass filter was performed to identify degradation in triclosan. Degradation can be concluded if there is a reduction in parent ion intensity or if there were other prominent peaks other than the parent ion peak.
  • the stability of the formulation was evaluated after 8 weeks of storage.
  • TCS-HPpCD-NaCl composition produced using the slurry method as described herein was weighed and dissolved in 10 ml MilliQ water. Following that, it was filtered using 0.22 ⁇ Millex® GP Filter Units. The stability of the composition was evaluated for 8 weeks under two conditions:
  • Example 35 Triclosan saturation in 2-hydroxypropyl-fi-cyclodextrin ( ⁇ ) with carboxymethyl cellulose (CMC)
  • HPpCD 2-hydroxypropyl-P-cyclodextrin
  • CMC carboxymethyl cellulose
  • Example 36 Minimum Inhibitory Concentration (MIC) of triclosan compositions- Macrodilution ( tube ) broth assay
  • MIC Minimum Inhibitory Concentration
  • the minimum inhibitory concentration (MIC) of the TCS-based formulation was found to be between 24 ng mL “1 to 30 ng mL "1 , from a 72-hour incubation period at 37°C.
  • the test samples were deliberately inoculated with a 24-hour broth culture of Staphylococcus aureus.
  • the minimum inhibitory concentration (MIC) of the TCS-based formulation with 1% w/v saline was found to be between 12 ng mL "1 to 24 ng mL "1 , after a 72-hour incubation period at 37°C.
  • the test samples were deliberately inoculated with a 24-hour broth culture of Staphylococcus aureus.
  • Example 37 Minimum Inhibitory Concentration (MIC) of triclosan compositions - Microdilution assay
  • MIC Minimum Inhibitory Concentration
  • TCS-HPpCD TCS-HPpCD-NaCl
  • MRSA 1, MRSA 2, MRSA 3, MRSA 7, Mu50, WIS and MSSA 2590 using the Minimum Inhibitory Concentration (MIC) Broth Microdilution Assay.
  • the assay was conducted as described previously. Out of the clinically-isolated MRSA strains, MRSA 1 and MRSA 2 are resistant to mupirocin. They are characterised by the presence of the mupA gene.
  • the minimum inhibitory concentration (MIC) of the triclosan-based formulation was found to be between 24 ng/ml to 30 ng/ml; and between 12 ng/ml to 24 ng/ml for the formulation with 1% w/v saline added. This suggests a possible additive/synergistic effect between saline and the TCS-HPpCD-CMC formulation.
  • the MIC of the TCS -based formulation was found to be between 1 ng/ml to 0.2 ⁇ g/ml for Methicillin-resistant Staphylococcus aureus (MRSA) and Methicillin-sensitive Staphylococcus aureus (MSSA) bacteria strains.
  • MIC24 Minimum Inhibitory Concentration (MIC) performed using the Broth Microdilution Method with 7 different strains of Staphylococcus aureus. The MIC at 24 hours (MIC24) for MRSA 2 and MRSA 3 for 3mg/ml TCS in 80mM HPpCD with 2.8% NaCl was unable to be conclusively determined. MIC24 of MRSA and MSSA strains are comparable.
  • Example 38 Time-dependent bacterial activity of triclosan compositions
  • the time-dependent bactericidal kill profile of two compositions (TCS-HPpCD, TCS-HPpCD-NaCl) at full-strength and half-strength were determined against Staphylococcus aureus strains: MRS A 1, MRS A 2, MRSA7 and Mu50.
  • the compositions were dissolved directly in Mueller Hinton (MH) broth in a 15 ml falcon tube.
  • Half-strength concentrations were prepared by performing a two-fold dilution of the full-strength concentrations.
  • Example 39 Ex-vivo swab culture test of triclosan compositions
  • TCS-HPpCD-CMC-NaCl composition (0.3% w/v TCS) prepared by dry mixing was evaluated for its efficacy as a nebulising solution against the nasal swab samples of 'healthy' volunteers.
  • a total of twenty volunteers were recruited for the study. These volunteers were screened preliminarily through an online sign-up process (Google Forms) and particulars were collated in accordance with the Personal Data Protection Act 2012.
  • the exclusion criteria for the study was: (1) A history of any nose operations or procedures (2) History of allergic rhinitis, sinusitis or nasal mucositis (3) Chronic use of any nasal products (4) Chronic use of any antibiotics through any administration route. Volunteers fulfilling these criteria were deemed to be 'healthy'.
  • Copan Innovation Sterile Cotton Swabs (150C Cotton; Lot: B25 GCGL00) were purchased for the study.
  • Volunteers were requested to perform hand-washing steps with 4% w/v chlorhexidine gluconate solution, and lather on their entire hand surface 4% w/v chlorhexidine gluconate gel. The hands were allowed to either air-dry or dried using an automated hand-dryer. The volunteers then placed on Biomedia Powder-free Latex Examination Gloves (Lot: 110420011300) and then disinfected the gloves with 70% ethanol. Under close supervision of the investigator, the volunteers were asked to carefully remove the sterile cotton swabs from the protective casings, taking care not to allow the swab to come into contact with any surfaces.
  • the swab was then inserted about two centimetres into one nostril, and rested against the nasal septum. Light pressure was applied on the outside of the nose against the swab, such that the swab was held steadily in place between the septum and mucosa membrane. The swab was rotated against the mucosa for a total of 5 revolutions lasting 1 minute. The swab was then placed back into the protective casing, and another swab was similarly performed for the other nostril.
  • the two swab samples collected from each volunteer were subjected to different procedures.
  • One swab was placed directly into 10 ml of a normal-strength nutrient broth solution; the other was subject to 2 ml of the nebulised solution for a total of 5 minutes.
  • the swab was clamped using a retort stand and aligned directly against the nebuliser (set-up as shown in Figure 25).
  • the swab was rotated 90° clockwise every 1 minute and 15 seconds to ensure maximal coverage of the swab by the nebulised solution.
  • the treated swab was then placed in 10 ml of normal-strength nutrient broth solution. Both tubes were incubated at 37°C for 24 hours up to 72 hours. The tubes were observed for turbidity, indicative of microbiological growth, in intervals of 24 hours up to 72 hours and were measured against McFarland Equivalence Standards (Batch No.: 1313947; TM4000T10). All apparatus (i.e. glass tubes, measuring cylinders, pipette tips, stock bottles, and volumetric flasks), nutrient broth solution and sterile water were placed through a moist heat sterilisation process (autoclave) at 121°C for 15 minutes before they were used.
  • autoclave moist heat sterilisation process
  • Table 27 Swab culture results from twenty 'healthy' volunteers. Two swabs were obtained from each subject; and one swab was treated with the TCS formulation using a nebuliser, the other swab was used as a control. There is a statistically significant difference (p-value ⁇ 0.001) between the 72 hour cell densities of the positive controls and tests. * Based on McFarland's Standards; readings are obtained after incubation at 37°C for 72

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Epidemiology (AREA)
  • Inorganic Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Organic Chemistry (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Otolaryngology (AREA)
  • Communicable Diseases (AREA)
  • Oncology (AREA)
  • Medicinal Preparation (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

La présente invention concerne des formulations comprenant des agents antimicrobiens hydrophobes et leurs utilisations. Plus précisément, la présente invention concerne une composition comprenant un agent antimicrobien hydrophobe et une cyclodextrine.
PCT/SG2015/050435 2014-11-07 2015-11-05 Formulations comprenant des agents antimicrobiens avec des parties hydrophobes et leurs utilisations WO2016072939A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
SG10201407358Q 2014-11-07
SG10201407358Q 2014-11-07

Publications (1)

Publication Number Publication Date
WO2016072939A1 true WO2016072939A1 (fr) 2016-05-12

Family

ID=55909518

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/SG2015/050435 WO2016072939A1 (fr) 2014-11-07 2015-11-05 Formulations comprenant des agents antimicrobiens avec des parties hydrophobes et leurs utilisations

Country Status (1)

Country Link
WO (1) WO2016072939A1 (fr)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2020228917A1 (fr) * 2019-05-14 2020-11-19 Coloplast A/S Compositions élastomères de silicone comprenant du glycérol, de la cyclodextrine et de l'octénidine
WO2022069478A1 (fr) * 2020-09-29 2022-04-07 Lighthouse Pharma GmbH Formes galénique anti-infectieuses destinées à produire un rinçage nasal

Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0306455A1 (fr) * 1987-08-31 1989-03-08 Warner-Lambert Company Complexes de cyclodextrine de composés bis-biguanido hexane
EP0803243A2 (fr) * 1996-04-24 1997-10-29 Pfizer Inc. Produits dentaires contenant des cyclodextrines et des composés phénoliques
WO1998056340A1 (fr) * 1997-06-09 1998-12-17 The Procter & Gamble Company Compositions parfumees et procedes de reduction des odeurs corporelles et de l'humidite excessive
US5874067A (en) * 1996-10-24 1999-02-23 The Procter & Gamble Company Methods for controlling environmental odors on the body
US5955454A (en) * 1993-03-26 1999-09-21 Adir Et Compagnie Nasal pharmaceutical composition containing a progestogen
WO1999055815A1 (fr) * 1998-04-27 1999-11-04 The Procter & Gamble Company Compositions de cyclodextrine non complexee ameliorees pour l'elimination des odeurs
WO2009125246A1 (fr) * 2008-04-07 2009-10-15 Technopharma Sa Préparations ophtalmiques stables
WO2010114541A1 (fr) * 2009-04-01 2010-10-07 Colgate-Palmolive Company Dentifrice désensibilisant

Patent Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0306455A1 (fr) * 1987-08-31 1989-03-08 Warner-Lambert Company Complexes de cyclodextrine de composés bis-biguanido hexane
US5955454A (en) * 1993-03-26 1999-09-21 Adir Et Compagnie Nasal pharmaceutical composition containing a progestogen
EP0803243A2 (fr) * 1996-04-24 1997-10-29 Pfizer Inc. Produits dentaires contenant des cyclodextrines et des composés phénoliques
US5874067A (en) * 1996-10-24 1999-02-23 The Procter & Gamble Company Methods for controlling environmental odors on the body
WO1998056340A1 (fr) * 1997-06-09 1998-12-17 The Procter & Gamble Company Compositions parfumees et procedes de reduction des odeurs corporelles et de l'humidite excessive
WO1999055815A1 (fr) * 1998-04-27 1999-11-04 The Procter & Gamble Company Compositions de cyclodextrine non complexee ameliorees pour l'elimination des odeurs
WO2009125246A1 (fr) * 2008-04-07 2009-10-15 Technopharma Sa Préparations ophtalmiques stables
WO2010114541A1 (fr) * 2009-04-01 2010-10-07 Colgate-Palmolive Company Dentifrice désensibilisant

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
"ALLIANCE FOR THE PRUDENT USE OF ANTIBIOTICS", TRICLOSAN, January 2011 (2011-01-01), pages 1 - 18, Retrieved from the Internet <URL:http://www.tufts.edu/med/apua/consumers/personal_home_21_4240495089.pdf> [retrieved on 20160122] *
ABDEL-HALIM, E. S. ET AL.: "Cotton fabric finished with beta-cyclodextrin: Inclusion ability toward antimicrobial agent", CARBOHYDRATE POLYMERS., vol. 102, 2014, pages 550 - 556 *
CERN, A ET AL.: "Effect of Solubilizing Agents on Mupirocin Loading into and Release from PEGylated Nanoliposomes", JOURNAL OF PHARMACEUTICAL SCIENCES., vol. 103, no. 7, July 2014 (2014-07-01), pages 2131 - 2138 *

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2020228917A1 (fr) * 2019-05-14 2020-11-19 Coloplast A/S Compositions élastomères de silicone comprenant du glycérol, de la cyclodextrine et de l'octénidine
CN113795518A (zh) * 2019-05-14 2021-12-14 科洛普拉斯特公司 包含甘油、环糊精和奥替尼啶的弹性体硅酮组合物
CN113795518B (zh) * 2019-05-14 2023-06-09 科洛普拉斯特公司 包含甘油、环糊精和奥替尼啶的弹性体硅酮组合物
WO2022069478A1 (fr) * 2020-09-29 2022-04-07 Lighthouse Pharma GmbH Formes galénique anti-infectieuses destinées à produire un rinçage nasal

Similar Documents

Publication Publication Date Title
CN102448496B (zh) 含碘和类固醇的药物组合物及其用于治疗鼻炎疾病的用途
EP2395985B1 (fr) Compositions antiseptiques renfermant des ions argent et du menthol et leurs utilisations
WO2018071810A1 (fr) Compositions antibiotiques pour irrigation nasale et procédés
JP6941889B2 (ja) インサイチュゲル形成医薬組成物および副鼻腔疾患のためのその使用
CA2646446A1 (fr) Compositions pharmaceutiques pour favoriser la cicatrisation
MX2011001812A (es) Composicion topica de hidrogel.
CA2795611C (fr) Creme medicinale a base d&#39;acide fusidique produite au moyen de fusidate de sodium et incorporant un biopolymere et son procede de fabrication
EP2410975A1 (fr) Crème médicinale antibactérienne et son procédé de production
WO2016072939A1 (fr) Formulations comprenant des agents antimicrobiens avec des parties hydrophobes et leurs utilisations
WO2010109423A1 (fr) Crème médicinale antifongique à base de stéroïdes et comprenant du chitosane, et son procédé de fabrication
CN115209954B (zh) 用于治疗呼吸系统病变的组合物
EP2985019B1 (fr) Composition nasale ayant des propriétés antivirales
CN104147603A (zh) 一种新型海藻提取物组合物在预防病毒性感冒中的应用
TW202227104A (zh) 預防感染之組合物
WO2010109418A1 (fr) Crème médicinale antifongique et son procédé de fabrication
RU2536266C2 (ru) Крем медицинского назначения, изготовленный с использованием фрамицетина сульфата и хитозана, и способ его изготовления
EP2985027B1 (fr) Composition nasale renfermant un mélange d&#39;acides hyaluroniques et une solution saline
EP3313377B1 (fr) Compositions et procédés d&#39;utilisation de corps lamellaires pour objectifs thérapeutiques
ES2749128T3 (es) Composición para el tratamiento de rinitis
WO2011027246A1 (fr) Crème contenant du nitrate de miconazole et biopolymère destiné au traitement de l&#39;érythème fessier du nourrisson
KR20230145034A (ko) 진균 감염의 치료 및/또는 예방용 조성물
WO2010122477A1 (fr) Crème stéroïde médicale antibactérienne et antifongique incorporant un biopolymère, et procédé de fabrication de celle-ci
WO2012049545A1 (fr) Crème médicinale à base d&#39;acide fusidique réalisée au moyen de fusidate de sodium et par incorporation d&#39;un biopolymère, procédé permettant de fabriquer une telle crème

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 15856998

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 15856998

Country of ref document: EP

Kind code of ref document: A1